Heterocycle-aryl compounds for inflammation and immune-related uses

ABSTRACT

The invention relates to compounds that are useful as immunosuppressive agents and for treating and preventing inflammatory conditions, allergic disorders, and immune disorders.

RELATED APPLICATIONS

This application is a national stage filing under 35 U.S.C.§371 ofInternational Application No. PCT/US2008/009305, filed Jul. 31, 2008,which claims priority under 35 U.S.C.§119(e) to U.S. ProvisionalApplication No. 60/962,824, filed Aug. 1, 2007. The contents of each ofthese applications are incorporated herein by reference in theirentireties for all purposes.

FIELD OF THE INVENTION

This invention relates to biologically active chemical compounds, namelyphenyl, pyrazinyl, and pyridinyl derivatives that may be used forimmunosuppression or to treat or prevent inflammatory conditions,allergic disorders and immune disorders.

BACKGROUND OF THE INVENTION

Inflammation is a mechanism that protects mammals from invadingpathogens. However, while transient inflammation is necessary to protecta mammal from infection, uncontrolled inflammation causes tissue damageand is the underlying cause of many illnesses. Inflammation is typicallyinitiated by binding of an antigen to T-cell antigen receptor. Antigenbinding by a T-cell initiates calcium influx into the cell via calciumion channels, such as Ca²⁺-release-activated Ca²⁺ channels (CRAC).Calcium ion influx in turn initiates a signaling cascade that leads toactivation of these cells and an inflammatory response characterized bycytokine production.

Interleukin 2 (IL-2) is a cytokine that is secreted by T cells inresponse to calcium ion influx into the cell. IL-2 modulatesimmunological effects on many cells of the immune system. For example,it is a potent T cell mitogen that is required for T cell proliferation,promoting their progression from G1 to S phase of the cell cycle; itstimulates the growth of NK cells; and it acts as a growth factor to Bcells and stimulates antibody synthesis.

IL-2, although useful in the immune response, can cause a variety ofproblems. IL-2 damages the blood-brain barrier and the endothelium ofbrain vessels. These effects may be the underlying causes ofneuropsychiatric side effects observed under IL-2 therapy, e.g. fatigue,disorientation and depression. It also alters the electrophysiologicalbehavior of neurons.

Due to its effects on both T and B cells, IL-2 is a major centralregulator of immune responses. It plays a role in inflammatoryreactions, tumor surveillance, and hematopoiesis. It also affects theproduction of other cytokines, inducing IL-1, TNF-α and TNF-β secretion,as well as stimulating the synthesis of IFN-γ in peripheral leukocytes.

T cells that are unable to produce IL-2 become inactive (anergic). Thisrenders them potentially inert to any antigenic stimulation they mightreceive in the future. As a result, agents which inhibit IL-2 productioncan be used for immunosuppression or to treat or prevent inflammationand immune disorders. This approach has been clinically validated withimmunosuppressive drugs such as cyclosporin, FK506, and RS61443. Despitethis proof of concept, agents that inhibit IL-2 production remain farfrom ideal. Among other problems, efficacy limitations and unwanted sideeffects (including dose-dependant nephrotoxicity and hypertension)hinder their use.

Over production of proinflammatory cytokines other than IL-2 has alsobeen implicated in many autoimmune diseases. For example, Interleukin 5(IL-5), a cytokine that increases the production of eosinophils, isincreased in asthma. Overproduction of IL-5 is associated withaccumulation of eosinophils in the asthmatic bronchial mucosa, a hallmark of allergic inflammation. Thus, patients with asthma and otherinflammatory disorders involving the accumulation of eosinophils wouldbenefit from the development of new drugs that inhibit the production ofIL-5.

Interleukin 4 (IL-4) and interleukin 13 (IL-13) have been identified asmediators of the hypercontractility of smooth muscle found ininflammatory bowel disease and asthma. Thus, patients with asthma andinflammatory bowel disease would benefit from the development of newdrugs that inhibit IL-4 and IL-13 production.

Granulocyte macrophage-colony stimulating factor (GM-CSF) is a regulatorof maturation of granulocyte and macrophage lineage population and hasbeen implicated as a key factor in inflammatory and autoimmune diseases.Anti-GM-CSF antibody blockade has been shown to ameliorate autoimmunedisease. Thus, development of new drugs that inhibit the production ofGM-CSF would be beneficial to patients with an inflammatory orautoimmune disease.

There is a continuing need for new drugs which overcome one or more ofthe shortcomings of drugs currently used for immunosuppression or in thetreatment or prevention of inflammatory disorders, allergic disordersand autoimmune disorders. Desirable properties of new drugs includeefficacy against diseases or disorders that are currently untreatable orpoorly treatable, new mechanism of action, oral bioavailability and/orreduced side effects.

SUMMARY OF THE INVENTION

This invention meets the above-mentioned needs by providing certainphenyl and pyridinyl derivatives that inhibit the activity of CRAC ionchannels and inhibit the production of IL-2, IL-4, IL-5, IL-13, GM-CSF,TNF-α, and IFNγ. These compounds are particularly useful forimmunosuppression and/or to treat or prevent inflammatory conditions,allergic disorders and immune disorders.

In one embodiment, the invention relates to compounds

-   2,6-difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(thiazol-2-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(pyridin-3-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(pyridin-3-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1,3,4-oxadiazol-2-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(6-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamide;-   N-(4-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(4-methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamide;-   N-(4-(5-(5-chloro-2-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   3-methyl-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamide;-   N-(4-(5-(4-chloropyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(5-(4-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide    hydrochloride; or-   N-(4-(5-(1-ethyl-1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;    or a pharmaceutically acceptable salt, solvate, clathrate, or    prodrug thereof.

A compound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof is particularly useful inhibitingimmune cell (e.g., T-cells and/or B-cells) activation (e.g., activationin response to an antigen). In particular, a compound of the inventionor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof can inhibit the production of certain cytokines that regulateimmune cell activation. For example, a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrug thereofcan inhibit the production of IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α,INF-γ or combinations thereof. Moreover, a compound of the invention ora pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof can modulate the activity of one or more ion channel involved inactivation of immune cells, such as CRAC ion channels.

In one embodiment, compounds of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof are particularlyuseful for inhibiting mast cell degranulation. Mast cell degranulationhas been implicated in allergic reactions.

A compound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof is particularly useful forimmunosuppression or for treating or preventing inflammatory conditions,allergic disorders, and immune disorders.

The invention also encompasses pharmaceutical compositions comprising acompound of the invention or a pharmaceutically acceptable salt,solvate, clathrate, or prodrug thereof; and a pharmaceuticallyacceptable carrier or vehicle. These compositions may further compriseadditional agents. These compositions are useful for immunosuppressionand treating or preventing inflammatory conditions, allergic disordersand immune disorders.

The invention further encompasses methods for treating or preventinginflammatory conditions, allergic disorders, and immune disorders,comprising administering to a subject in need thereof an effectiveamount of a compound of the invention or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof, or a pharmaceuticalcomposition comprising a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof. These methodsmay also comprise administering to the subject an additional agentseparately or in a combination composition with the compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof.

The invention further encompasses methods for suppressing the immunesystem of a subject, comprising administering to a subject in needthereof an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof, or a pharmaceutical composition comprising a compound of theinvention or a pharmaceutically acceptable salt, solvate, clathrate, orprodrug thereof. These methods may also comprise administering to thesubject an additional agent separately or in a combination compositionwith the compound of the invention or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof.

The invention further encompasses methods for inhibiting immune cellactivation, including inhibiting proliferation of T cells and/or Bcells, in vivo or in vitro comprising administering to the cell aneffective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof or apharmaceutical composition comprising a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

The invention further encompasses methods for inhibiting cytokineproduction in a cell (e.g., IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α,and/or INF-γ production) in vivo or in vitro comprising administering toa cell an effective amount of a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrug thereofor a pharmaceutical composition comprising a compound of the inventionor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

The invention further encompasses methods for modulating ion channelactivity (e.g., CRAC) in vivo or in vitro comprising administering aneffective amount of a compound of the invention or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof or apharmaceutical composition comprising a compound of the invention or apharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

All of the methods of this invention may be practice with a compound ofthe invention alone, or in combination with other agents, such as otherimmunosuppressive agents, anti-inflammatory agents, agents for thetreatment of allergic disorders or agents for the treatment of immunedisorders.

DETAILED DESCRIPTION OF THE INVENTION Definitions

Unless otherwise specified, the below terms used herein are defined asfollows:

As used herein, the term an “aromatic ring” or “aryl” means a monocyclicor polycyclic-aromatic ring or ring radical comprising carbon andhydrogen atoms. Examples of suitable aryl groups include, but are notlimited to, phenyl, tolyl, anthacenyl, fluorenyl, indenyl, azulenyl, andnaphthyl, as well as benzo-fused carbocyclic moieties such as5,6,7,8-tetrahydronaphthyl. An aryl group can be unsubstituted orsubstituted with one or more substituents (including without limitationalkyl (preferably, lower alkyl or alkyl substituted with one or morehalo), hydroxy, alkoxy (preferably, lower alkoxy), alkylsulfanyl, cyano,halo, amino, and nitro. In certain embodiments, the aryl group is amonocyclic ring, wherein the ring comprises 6 carbon atoms.

As used herein, the term “alkyl” means a saturated straight chain orbranched non-cyclic hydrocarbon typically having from 1 to 10 carbonatoms. Representative saturated straight chain alkyls include methyl,ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyland n-decyl; while saturated branched alkyls include isopropyl,sec-butyl, isobutyl, tert-butyl, isopentyl, 2-methylbutyl,3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl,2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl,2,3-dimethylbutyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl,2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl,2,2-dimethylpentyl, 2,2-dimethylhexyl, 3,3-dimethylpentyl,3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylpentyl, 3-ethylpentyl,2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl,2-methyl-3-ethylpentyl, 2-methyl-4-ethylpentyl, 2-methyl-2-ethylhexyl,2-methyl-3-ethylhexyl, 2-methyl-4-ethylhexyl, 2,2-diethylpentyl,3,3-diethylhexyl, 2,2-diethylhexyl, 3,3-diethylhexyl and the like. Alkylgroups included in compounds of this invention may be optionallysubstituted with one or more substituents. Examples of substituentsinclude, but are not limited to, amino, alkylamino, alkoxy,alkylsulfanyl, oxo, halo, acyl, nitro, hydroxyl, cyano, aryl, alkylaryl,aryloxy, arylsulfanyl, arylamino, carbocyclyl, carbocyclyloxy,carbocyclylthio, carbocyclylamino, heterocyclyl, heterocyclyloxy,heterocyclylamino, heterocyclylthio, and the like. In addition, anycarbon in the alkyl segment may be substituted with oxygen (═O), sulfur(═S), or nitrogen (═NR²², wherein R²² is —H, an alkyl, acetyl, oraralkyl). Lower alkyls are typically preferred for the compounds of thisinvention.

The term alkylene refers to an alkyl group or a cycloalkyl group thathas two points of attachment to two moieties (e.g., {—CH₂—}, —{CH₂CH₂—},

etc., wherein the brackets indicate the points of attachment). Alkylenegroups may be substituted or unsubstituted with one or moresubstituents.

An aralkyl group refers to an aryl group that is attached to anothermoiety via an alkylene linker. Aralkyl groups can be substituted orunsubstituted with one or more substituents.

The term “alkoxy,” as used herein, refers to an alkyl group which islinked to another moiety though an oxygen atom. Alkoxy groups can besubstituted or unsubstituted with one or more substituents.

The term “alkylsulfanyl,” as used herein, refers to an alkyl group whichis linked to another moiety though a divalent sulfur atom. Alkylsulfanylgroups can be substituted or unsubstituted with one or moresubstituents.

The term “arylsulfanyl,” as used herein, refers to an aryl group whichis linked to another moiety though a divalent sulfur atom. Arylsulfanylgroups can be substituted or unsubstituted with one or moresubstituents.

The term “alkyl ester” as used herein, refers to a group represented bythe formula —C(O)OR₃₂, wherein R₃₂ is an alkyl group. A lower alkylester is a group represented by the formula —C(O)OR₃₂, wherein R₃₂ is alower alkyl group.

The term “heteroalkyl,” as used herein, refers to an alkyl group whichhas one or more carbons in the alkyl chain replaced with an —O—, —S— or—NR₂₇—, wherein R₂₇ is H or a lower alkyl. Heteroalkyl groups can besubstituted or unsubstituted with one or more substituents.

The term “alkylamino,” as used herein, refers to an amino group in whichone hydrogen atom attached to the nitrogen has been replaced by an alkylgroup. The term “dialkylamino,” as used herein, refers to an amino groupin which two hydrogen atoms attached to the nitrogen have been replacedby alkyl groups, in which the alkyl groups can be the same or different.Alkylamino groups and dialkylamino groups can be substituted orunsubstituted with one or more substituents.

As used herein, the term “alkenyl” means a straight chain or branched,hydrocarbon radical typically having from 2 to 10 carbon atoms andhaving at least one carbon-carbon double bond. Representative straightchain and branched alkenyls include vinyl, allyl, 1-butenyl, 2-butenyl,isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,1-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, 1-hexenyl, 2-hexenyl,3-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 1-octenyl, 2-octenyl,3-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl, 1-decenyl, 2-decenyl,3-decenyl and the like. Alkenyl groups can be substituted orunsubstituted with one or more substituents.

As used herein, the term “alkynyl” means a straight chain or branched,hydrocarbonon radical typically having from 2 to 10 carbon atoms andhaving at lease one carbon-carbon triple bond. Representative straightchain and branched alkynyls include acetylenyl, propynyl, 1-butynyl,2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, 4-pentynyl,-1-hexynyl, 2-hexynyl, 5-hexynyl, 1-heptynyl, 2-heptynyl, 6-heptynyl,1-octynyl, 2-octynyl, 7-octynyl, 1-nonynyl, 2-nonynyl, 8-nonynyl,1-decynyl, 2-decynyl, 9-decynyl and the like. Alkynyl groups can besubstituted or unsubstituted with one or more substituents.

As used herein, the term “cycloalkyl” means a saturated, mono- orpolycyclic alkyl radical typically having from 3 to 14 carbon atoms.Representative cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantly,decahydronaphthyl, octahydropentalene, bicycle[1.1.1]pentanyl, and thelike. Cycloalkyl groups can be substituted or unsubstituted with one ormore substituents.

As used herein, the term “cycloalkenyl” means a cyclic non-aromaticalkenyl radical having at least one carbon-carbon double bond in thecyclic system and typically having from 5 to 14 carbon atoms.Representative cycloalkenyls include cyclopentenyl, cyclopentadienyl,cyclohexenyl, cyclohexadienyl, cycloheptenyl, cycloheptadienyl,cycloheptatrienyl, cyclooctenyl, cyclooctadienyl, cyclooctatrienyl,cyclooctatetraenyl, cyclononenyl, cyclononadienyl, cyclodecenyl,cyclodecadienyl and the like. Cycloalkenyl groups can be substituted orunsubstituted with one or more substituents.

As used herein, the term “heterocycle” or “heterocyclyl” means amonocyclic or polycyclic heterocyclic ring (typically having 3- to14-members) which is either a saturated ring or an unsaturatednon-aromatic ring. A 3-membered heterocycle can contain up to 3heteroatoms, and a 4- to 14-membered heterocycle can contain from 1 toabout 8 heteroatoms. Each heteroatom is independently selected fromnitrogen, which can be quaternized; oxygen; and sulfur, includingsulfoxide and sulfone. The heterocycle may be attached via anyheteroatom or carbon atom. Representative heterocycles includemorpholinyl, thiomorpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl,piperazinyl, hydantoinyl, valerolactamyl, oxiranyl, oxetanyl,tetrahydrofuranyl, tetrahydropyranyl, 4H-pyranyl, tetrahydropyrindinyl,tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, andthe like. A heteroatom may be substituted with a protecting group knownto those of ordinary skill in the art, for example, the hydrogen on anitrogen may be substituted with a tert-butoxycarbonyl group.Furthermore, the heterocyclyl may be optionally substituted with one ormore substituents (including without limitation a halo, an alkyl, ahaloalkyl, or aryl). Only stable isomers of such substitutedheterocyclic groups are contemplated in this definition.

As used herein, the term “heteroaromatic” or “heteroaryl” means amonocyclic or polycyclic heteroaromatic ring (or radical thereof)comprising carbon atom ring members and one or more heteroatom ringmembers (such as, for example, oxygen, sulfur or nitrogen). Typically,the heteroaromatic ring has from 5 to about 14 ring members in which atleast 1 ring member is a heteroatom selected from oxygen, sulfur andnitrogen. In another embodiment, the heteroaromatic ring is a 5 or 6membered ring and may contain from 1 to about 4 heteroatoms. In anotherembodiment, the heteroaromatic ring system has a 7 to 14 ring membersand may contain from 1 to about 7 heteroatoms. Representativeheteroaryls include pyridyl, furyl, thienyl, pyrrolyl, oxazolyl,imidazolyl, indolizinyl, thiazolyl, isoxazolyl, pyrazolyl, isothiazolyl,pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, triazolyl, pyridinyl,thiadiazolyl, pyrazinyl, quinolyl, isoquniolyl, indazolyl, benzoxazolyl,benzofuryl, benzothiazolyl, indolizinyl, imidazopyridinyl, isothiazolyl,tetrazolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl,benzothiadiazolyl, benzoxadiazolyl, indolyl, tetrahydroindolyl,azaindolyl, imidazopyridyl, qunizaolinyl, purinyl,pyrrolo[2,3]pyrimidyl, pyrazolo[3,4]pyrimidyl or benzo(b)thienyl and thelike. Heteroaryl groups may be optionally substituted with one or moresubstituents

A heteroaralkyl group refers to a heteroaryl group that is attached toanother moiety via an alkylene linker. Heteroaralkyl groups can besubstituted or unsubstituted with one or more substituents.

As used herein, the term “halogen” or “halo” means —F, —Cl, —Br or —I.

As used herein, the term “haloalkyl” means an alkyl group in which oneor more —H is replaced with a halo group. Examples of haloalkyl groupsinclude —CF₃, —CHF₂, —CCl₃, —CH₂CH₂Br, —CH₂CH(CH₂CH₂Br)CH₃, —CHICH₃, andthe like.

As used herein, the term “haloalkoxy” means an alkoxy group in which oneor more —H is replaced with a halo group. Examples of haloalkoxy groupsinclude —OCF₃ and —OCHF₂.

The terms “bioisostere” and “bioisosteric replacement” have the samemeanings as those generally recognized in the art. Bioisosteres areatoms, ions, or molecules in which the peripheral layers of electronscan be considered substantially identical. The term bioisostere isusually used to mean a portion of an overall molecule, as opposed to theentire molecule itself. Bioisosteric replacement involves using onebioisostere to replace another with the expectation of maintaining orslightly modifying the biological activity of the first bioisostere. Thebioisosteres in this case are thus atoms or groups of atoms havingsimilar size, shape and electron density. Preferred bioisosteres ofesters, amides or carboxylic acids are compounds containing two sitesfor hydrogen bond acceptance. In one embodiment, the ester, amide orcarboxylic acid bioisostere is a 5-membered monocyclic heteroaryl ring,such as an optionally substituted 1H-imidazolyl, an optionallysubstituted oxazolyl, 1H-tetrazolyl, [1,2,4]triazolyl, or an optionallysubstituted [1,2,4]oxadiazolyl.

As used herein, the terms “subject”, “patient” and “animal”, are usedinterchangeably and include, but are not limited to, a cow, monkey,horse, sheep, pig, mini pig, chicken, turkey, quail, cat, dog, mouse,rat, rabbit, guinea pig and human. The preferred subject, patient oranimal is a human.

As used herein, the term “lower” refers to a group having up to fourcarbon atoms. For example, a “lower alkyl” refers to an alkyl radicalhaving from 1 to 4 carbon atoms, and a “lower alkenyl” or “loweralkynyl” refers to an alkenyl or alkynyl radical having from 2 to 4carbon atoms, respectively. A lower alkoxy or a lower alkylsulfanylrefers to an alkoxy or an alkylsulfanyl having from 1 to 4 carbon atoms.Lower substituents are typically preferred.

Where a particular substituent, such as an alkyl substituent, occursmultiple times in a given structure or moeity, the identity of thesubstituent is independent in each case and may be the same as ordifferent from other occurrences of that substituent in the structure ormoiety. Furthermore, individual substituents in the specific embodimentsand exemplary compounds of this invention are preferred in combinationwith other such substituents in the compounds of this invention, even ifsuch individual substituents are not expressly noted as being preferredor not expressly shown in combination with other substituents.

The compounds of the invention are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

Suitable substituents for an alkyl, alkoxy, alkylsulfanyl, alkylamino,dialkylamino, alkylene, alkenyl, alkynyl, cycloalkyl, cycloalkenyl,heterocyclyl, aryl, aralkyl, heteroaryl, and heteroaralkyl groupsinclude any substituent which will form a stable compound of theinvention. Examples of substituents for an alkyl, alkoxy, alkylsulfanyl,alkylamino, dialkylamino, alkylene, alkenyl, alkynyl, cycloalkyl,cycloalkenyl, heterocyclyl, aryl, aralkyl, heteroaryl, and heteroaralkylinclude an alkyl, an alkoxy, an alkylsulfanyl, an alkylamino, adialkylamino, an alkenyl, an alkynyl, a cycloalkyl, a cycloalkenyl, aheterocyclyl, an aryl, a heteroaryl, an aralkyl, a heteraralkyl, ahaloalkyl, —C(O)NR₂₃R₂₄, —NR₂₅C(O)R₂₆, halo, —OR₂₅, cyano, nitro,

haloalkoxy, —C(O)R₂₅, —NR₂₃R₂₄, —SR₂₅, —C(O)OR₂₅, —OC(O)R₂₅,—NR₂₅C(O)NR₂₃R₂₄, —OC (O)NR₂₃R₂₄, —NR₂₅C(O)OR₂₆, —S(O)_(p)R₂₅, or—S(O)_(p)NR₂₃R₂₄, wherein R₂₃ and R₂₄, for each occurrence are,independently, H, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, acycloalkenyl, a heterocyclyl, an aryl, a heteroaryl, an aralkyl, or aheteraralkyl; or R₂₃ and R₂₄ taken together with the nitrogen to whichthey are attached is a heterocyclyl or a heteroaryl; and R₂₅ and R₂₆ foreach occurrence are, independently, H, an alkyl, an alkenyl, an alkynyl,a cycloalkyl, a cycloalkenyl, a heterocyclyl, an aryl, a heteroaryl, anaralkyl, or a heteraralkyl;

In addition, alkyl, cycloalkyl, alkylene, a heterocyclyl, and anysaturated portion of a alkenyl, cycloalkenyl, alkynyl, aralkyl, andheteroaralkyl groups, may also be substituted with ═O, ═S, ═N—R₂₂.

When a heterocyclyl, heteroaryl, or heteroaralkyl group contains anitrogen atom, it may be substituted or unsubstituted. When a nitrogenatom in the aromatic ring of a heteroaryl group has a substituent thenitrogen may be a quaternary nitrogen.

Choices and combinations of substituents and variables envisioned bythis invention are only those that result in the formation of stablecompounds. The term “stable”, as used herein, refers to compounds whichpossess stability sufficient to allow manufacture and which maintainsthe integrity of the compound for a sufficient period of time to beuseful for the purposes detailed herein (e.g., therapeutic orprophylactic administration to a subject). Typically, such compounds arestable at a temperature of 40° C. or less, in the absence of excessivemoisture, for at least one week. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

Unless indicated otherwise, the compounds of the invention containingreactive functional groups (such as, without limitation, carboxy,hydroxy, and amino moieties) also include protected derivatives thereof.“Protected derivatives” are those compounds in which a reactive site orsites are blocked with one or more protecting groups. Suitableprotecting groups for carboxy moieties include benzyl, tert-butyl, andthe like. Suitable protecting groups for amino and amido groups includeacetyl, tert-butoxycarbonyl, benzyloxycarbonyl, and the like. Suitableprotecting groups for hydroxy include benzyl, trimethyl silyl (TMS) andthe like. Other suitable protecting groups are well known to those ofordinary skill in the art and include those found in T. W. Greene,Protecting Groups in Organic Synthesis, John Wiley & Sons, Inc. 1981,the entire teachings of which are incorporated herein by reference.

As used herein, the term “compound(s) of this invention” and similarterms refers to a compound of Table 1, or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof and also include protectedderivatives thereof.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide acompound of this invention. Prodrugs may only become active upon suchreaction under biological conditions, but they may have activity intheir unreacted forms. Examples of prodrugs contemplated in thisinvention include, but are not limited to, analogs or derivatives ofcompounds of Table 1 that comprise biohydrolyzable moieties such asbiohydrolyzable amides, biohydrolyzable esters, biohydrolyzablecarbamates, biohydrolyzable carbonates, biohydrolyzable ureides, andbiohydrolyzable phosphate analogues. Other examples of prodrugs includederivatives of compounds of Table 1 that comprise —NO, —NO₂, —ONO, or—ONO₂ moieties. Prodrugs can typically be prepared using well-knownmethods, such as those described by 1 BURGER'S MEDICINAL CHEMISTRY ANDDRUG DISCOVERY (1995) 172-178, 949-982 (Manfred E. Wolff ed., 5^(th)ed), the entire teachings of which are incorporated herein by reference.

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide”, “biohydrolyzable ester”, “biohydrolyzablecarbamate”, “biohydrolyzable carbonate”, “biohydrolyzable ureide” and“biohydrolyzable phosphate analogue” mean an amide, ester, carbamate,carbonate, ureide, or phosphate analogue, respectively, that either: 1)does not destroy the biological activity of the compound and confersupon that compound advantageous properties in vivo, such as uptake,duration of action, or onset of action; or 2) is itself biologicallyinactive but is converted in vivo to a biologically active compound.Examples of biohydrolyzable amides include, but are not limited to,lower alkyl amides, α-amino acid amides, alkoxyacyl amides, andalkylaminoalkylcarbonyl amides. Examples of biohydrolyzable estersinclude, but are not limited to, lower alkyl esters, alkoxyacyloxyesters, alkyl acylamino alkyl esters, and choline esters. Examples ofbiohydrolyzable carbamates include, but are not limited to, loweralkylamines, substituted ethylenediamines, aminoacids,hydroxyalkylamines, heterocyclic and heteroaromatic amines, andpolyether amines.

As used herein, the term “pharmaceutically acceptable salt,” is a saltformed from an acid and a basic group of one of the compounds ofTable 1. Illustrative salts include, but are not limited, to sulfate,citrate, acetate, oxalate, chloride, bromide, iodide, nitrate,bisulfate, phosphate, acid phosphate, isonicotinate, lactate,salicylate, acid citrate, tartrate, oleate, tannate, pantothenate,bitartrate, ascorbate, succinate, maleate, gentisinate, fumarate,gluconate, glucaronate, saccharate, formate, benzoate, glutamate,methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate,and pamoate (i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts.The term “pharmaceutically acceptable salt” also refers to a saltprepared from a compound of Table 1 having an acidic functional group,such as a carboxylic acid functional group, and a pharmaceuticallyacceptable inorganic or organic base. Suitable bases include, but arenot limited to, hydroxides of alkali metals such as sodium, potassium,and lithium; hydroxides of alkaline earth metal such as calcium andmagnesium; hydroxides of other metals, such as aluminum and zinc;ammonia, and organic amines, such as unsubstituted orhydroxy-substituted mono-, di-, or trialkylamines; dicyclohexylamine;tributyl amine; pyridine; N-methyl,N-ethylamine; diethylamine;triethylamine; mono-, bis-, or tris-(2-hydroxy-lower alkyl amines), suchas mono-, bis-, or tris-(2-hydroxyethyl)-amine,2-hydroxy-tert-butylamine, or tris-(hydroxymethyl)methylamine,N,N,-di-lower alkyl-N-(hydroxy lower alkyl)-amines, such asN,N-dimethyl-N-(2-hydroxyethyl)-amine, or tri-(2-hydroxyethyl)amine;N-methyl-D-glucamine; and amino acids such as arginine, lysine, and thelike. The term “pharmaceutically acceptable salt” also refers to a saltprepared from a compound of Table 1 having a basic functional group,such as an amino functional group, and a pharmaceutically acceptableinorganic or organic acid. Suitable acids include, but are not limitedto, hydrogen sulfate, citric acid, acetic acid, oxalic acid,hydrochloric acid, hydrogen bromide, hydrogen iodide, nitric acid,phosphoric acid, isonicotinic acid, lactic acid, salicylic acid,tartaric acid, ascorbic acid, succinic acid, maleic acid, besylic acid,fumaric acid, gluconic acid, glucaronic acid, saccharic acid, formicacid, benzoic acid, glutamic acid, methanesulfonic acid, ethanesulfonicacid, benzenesulfonic acid, and p-toluenesulfonic acid.

When a disclosed compound is named or depicted by structure, it is to beunderstood that solvates (e.g., hydrates) of the compound or itspharmaceutically acceptable salts are also included. “Solvates” refer tocrystalline forms wherein solvent molecules are incorporated into thecrystal lattice during crystallization. Solvate may include water ornonaqueous solvents such as ethanol, isopropanol, DMSO, acetic acid,ethanolamine, and EtOAc. Solvates, wherein water is the solvent moleculeincorporated into the crystal lattice, are typically referred to as“hydrates”. Hydrates include a stoichiometric or non-stoichiometricamount of water bound by non-covalent intermolecular forces.

When a disclosed compound is named or depicted by structure, it is to beunderstood that the compound, including solvates thereof, may exist incrystalline forms, non-crystalline forms or a mixture thereof. Thecompounds or solvates may also exhibit polymorphism (i.e. the capacityto occur in different crystalline forms). These different crystallineforms are typically known as “polymorphs.” It is to be understood thatwhen named or depicted by structure, the disclosed compounds andsolvates (e.g., hydrates) also include all polymorphs thereof. As usedherein, the term “polymorph” means solid crystalline forms of a compoundof the present invention or complex thereof. Different polymorphs of thesame compound can exhibit different physical, chemical and/orspectroscopic properties. Different physical properties include, but arenot limited to stability (e.g., to heat or light), compressibility anddensity (important in formulation and product manufacturing), anddissolution rates (which can affect bioavailability). Differences instability can result from changes in chemical reactivity (e.g.,differential oxidation, such that a dosage form discolors more rapidlywhen comprised of one polymorph than when comprised of anotherpolymorph) or mechanical characteristics (e.g., tablets crumble onstorage as a kinetically favored polymorph converts to thermodynamicallymore stable polymorph) or both (e.g., tablets of one polymorph are moresusceptible to breakdown at high humidity). Different physicalproperties of polymorphs can affect their processing. For example, onepolymorph might be more likely to form solvates or might be moredifficult to filter or wash free of impurities than another due to, forexample, the shape or size distribution of particles of it. In addition,one polymorph may spontaneously convert to another polymorph undercertain conditions.

When a disclosed compound is named or depicted by structure, it is to beunderstood that clathrates (“inclusion compounds”) of the compound orits pharmaceutically acceptable salts, solvates or polymorphs are alsoincluded. As used herein, the term “clathrate” means a compound of thepresent invention or a salt thereof in the form of a crystal latticethat contains spaces (e.g., channels) that have a guest molecule (e.g.,a solvent or water) trapped within.

As used herein, the term “asthma” means a pulmonary disease, disorder orcondition characterized by reversible airway obstruction, airwayinflammation, and increased airway responsiveness to a variety ofstimuli.

“Immunosuppression” refers to impairment of any component of the immunesystem resulting in decreased immune function. This impairment may bemeasured by any conventional means including whole blood assays oflymphocyte function, detection of lymphocyte proliferation andassessment of the expression of T cell surface antigens. The antisheepred blood cell (SRBC) primary (IgM) antibody response assay (usuallyreferred to as the plaque assay) is one specific method. This and othermethods are described in Luster, M. I., Portier, C., Pait, D. G., White,K. L., Jr., Gennings, C., Munson, A. E., and Rosenthal, G. J. (1992).“Risk Assessment in Immunotoxicology I: Sensitivity and Predictabilityof Immune Tests.” Fundam. Appl. Toxicol., 18, 200-210. Measuring theimmune response to a T-cell dependent immunogen is another particularlyuseful assay (Dean, J. H., House, R. V., and Luster, M. I. (2001).“Immunotoxicology: Effects of, and Responses to, Drugs and Chemicals.”In Principles and Methods of Toxicology: Fourth Edition (A. W. Hayes,Ed.), pp. 1415-1450, Taylor & Francis, Philadelphia, Pa.).

The compounds of this invention can be used to treat subjects withimmune disorders. As used herein, the term “immune disorder” and liketerms means a disease, disorder or condition caused by the immune systemof an animal, including autoimmune disorders. Immune disorders includethose diseases, disorders or conditions that have an immune componentand those that are substantially or entirely immune system-mediated.Autoimmune disorders are those wherein the animal's own immune systemmistakenly attacks itself, thereby targeting the cells, tissues, and/ororgans of the animal's own body. For example, the autoimmune reaction isdirected against the nervous system in multiple sclerosis and the gut inCrohn's disease. In other autoimmune disorders such as systemic lupuserythematosus (lupus), affected tissues and organs may vary amongindividuals with the same disease. One person with lupus may haveaffected skin and joints whereas another may have affected skin, kidney,and lungs. Ultimately, damage to certain tissues by the immune systemmay be permanent, as with destruction of insulin-producing cells of thepancreas in Type 1 diabetes mellitus. Specific autoimmune disorders thatmay be ameliorated using the compounds and methods of this inventioninclude without limitation, autoimmune disorders of the nervous system(e.g., multiple sclerosis, myasthenia gravis, autoimmune neuropathiessuch as Guillain-Barré, and autoimmune uveitis), autoimmune disorders ofthe blood (e.g., autoimmune hemolytic anemia, pernicious anemia, andautoimmune thrombocytopenia), autoimmune disorders of the blood vessels(e.g., temporal arteritis, anti-phospholipid syndrome, vasculitides suchas Wegener's granulomatosis, and Behcet's disease), autoimmune disordersof the skin (e.g., psoriasis, dermatitis herpetiformis, pemphigusvulgaris, and vitiligo), autoimmune disorders of the gastrointestinalsystem (e.g., Crohn's disease, ulcerative colitis, primary biliarycirrhosis, and autoimmune hepatitis), autoimmune disorders of theendocrine glands (e.g., Type 1 or immune-mediated diabetes mellitus,Grave's disease. Hashimoto's thyroiditis, autoimmune oophoritis andorchitis, and autoimmune disorder of the adrenal gland); and autoimmunedisorders of multiple organs (including connective tissue andmusculoskeletal system diseases) (e.g., rheumatoid arthritis, systemiclupus erythematosus, scleroderma, polymyositis, dermatomyositis,spondyloarthropathies such as ankylosing spondylitis, and Sjogren'ssyndrome). In addition, other immune system mediated diseases, such asgraft-versus-host disease and allergic disorders, are also included inthe definition of immune disorders herein. Because a number of immunedisorders are caused by inflammation, there is some overlap betweendisorders that are considered immune disorders and inflammatorydisorders. For the purpose of this invention, in the case of such anoverlapping disorder, it may be considered either an immune disorder oran inflammatory disorder. “Treatment of an immune disorder” hereinrefers to administering a compound or a composition of the invention toa subject, who has an immune disorder, a symptom of such a disease or apredisposition towards such a disease, with the purpose to cure,relieve, alter, affect, or prevent the autoimmune disorder, the symptomof it, or the predisposition towards it.

As used herein, the term “allergic disorder” means a disease, conditionor disorder associated with an allergic response against normallyinnocuous substances. These substances may be found in the environment(such as indoor air pollutants and aeroallergens) or they may benon-environmental (such as those causing dermatological or foodallergies). Allergens can enter the body through a number of routes,including by inhalation, ingestion, contact with the skin or injection(including by insect sting). Many allergic disorders are linked toatopy, a predisposition to generate the allergic antibody IgE. BecauseIgE is able to sensitize mast cells anywhere in the body, atopicindividuals often express disease in more than one organ.

For the purpose of this invention, allergic disorders include anyhypersensitivity that occurs upon re-exposure to the sensitizingallergen, which in turn causes the release of inflammatory mediators.Allergic disorders include without limitation, allergic rhinitis (e.g.,hay fever), sinusitis, rhinosinusitis, chronic or recurrent otitismedia, drug reactions, insect sting reactions, latex reactions,conjunctivitis, urticaria, anaphylaxis and anaphylactoid reactions,atopic dermatitis, asthma and food allergies.

The compounds of this invention can be used to prevent or to treatsubjects with inflammatory disorders. As used herein, an “inflammatorydisorder” means a disease, disorder or condition characterized byinflammation of body tissue or having an inflammatory component. Theseinclude local inflammatory responses and systemic inflammation. Examplesof such inflammatory disorders include: transplant rejection, includingskin graft rejection; chronic inflammatory disorders of the joints,including arthritis, rheumatoid arthritis, osteoarthritis and bonediseases associated with increased bone resorption; inflammatory boweldiseases such as ileitis, ulcerative colitis, Barrett's syndrome, andCrohn's disease; inflammatory lung disorders such as asthma, adultrespiratory distress syndrome, and chronic obstructive airway disease;inflammatory disorders of the eye including corneal dystrophy, trachoma,onchocerciasis, uveitis, sympathetic ophthalmitis and endophthalmitis;chronic inflammatory disorders of the gums, including gingivitis andperiodontitis; tuberculosis; leprosy; inflammatory diseases of thekidney including uremic complications, glomerulonephritis and nephrosis;inflammatory disorders of the skin including sclerodermatitis, psoriasisand eczema; inflammatory diseases of the central nervous system,including chronic demyelinating diseases of the nervous system, multiplesclerosis, AIDS-related neurodegeneration and Alzheimer's disease,infectious meningitis, encephalomyelitis, Parkinson's disease,Huntington's disease, amyotrophic lateral sclerosis and viral orautoimmune encephalitis; autoimmune disorders, immune-complexvasculitis, systemic lupus and erythematodes; systemic lupuserythematosus (SLE); and inflammatory diseases of the heart such ascardiomyopathy, ischemic heart disease hypercholesterolemia,atherosclerosis; as well as various other diseases with significantinflammatory components, including preeclampsia; chronic liver failure,brain and spinal cord trauma, and cancer. There may also be a systemicinflammation of the body, exemplified by gram-positive or gram negativeshock, hemorrhagic or anaphylactic shock, or shock induced by cancerchemotherapy in response to pro-inflammatory cytokines, e.g., shockassociated with pro-inflammatory cytokines. Such shock can be induced,e.g., by a chemotherapeutic agent used in cancer chemotherapy.“Treatment of an inflammatory disorder” herein refers to administering acompound or a composition of the invention to a subject, who has aninflammatory disorder, a symptom of such a disorder or a predispositiontowards such a disorder, with the purpose to cure, relieve, alter,affect, or prevent the inflammatory disorder, the symptom of it, or thepredisposition towards it.

An “effective amount” is the quantity of compound in which a beneficialoutcome is achieved when the compound is administered to a subject oralternatively, the quantity of compound that possess a desired activityin-vivo or in-vitro. In the case of inflammatory disorders andautoimmune disorders, a beneficial clinical outcome includes reductionin the extent or severity of the symptoms associated with the disease ordisorder and/or an increase in the longevity and/or quality of life ofthe subject compared with the absence of the treatment. The preciseamount of compound administered to a subject will depend on the type andseverity of the disease or condition and on the characteristics of thesubject, such as general health, age, sex, body weight and tolerance todrugs. It will also depend on the degree, severity and type ofinflammatory disorder, autoimmune disorder, allergic disorder, or thedegree of immunosuppression sought. The skilled artisan will be able todetermine appropriate dosages depending on these and other factors.Effective amounts of the disclosed compounds typically range betweenabout 1 mg/m² per day and about 10 grams/m² per day, and preferablybetween 10 mg/m² per day and about 1 gram/m².

The compounds of the invention may contain one or more chiral centersand/or double bonds and, therefore, may exist as stereoisomers, such asdouble-bond isomers (i.e., geometric isomers), enantiomers, ordiastereomers. According to this invention, the chemical structuresdepicted herein, including the compounds of this invention, encompassall of the corresponding compounds' enantiomers and stereoisomers, thatis, both the stereomerically pure form (e.g., geometrically pure,enantiomerically pure, or diastereomerically pure) and enantiomeric,diastereomeric, and geometric isomeric mixtures. In some cases, oneenantiomer, diastereomer, or geometric isomer will possess superioractivity or an improved toxicity or kinetic profile compared to others.In those cases, such enantiomers, diastereomers, and geometric isomersof a compound of this invention are preferred.

The term “inhibit production of IL-2” and like terms means inhibitingIL-2 synthesis (e.g. by inhibiting transcription (mRNA expression), ortranslation (protein expression)) and/or inhibiting IL-2 secretion in acell that has the ability to produce and/or secrete IL-2 (e.g., Tlymphocyte). Likewise, the term “inhibiting production of IL-4, IL-5,IL-13, GM-CSF, TNF-α or INF-γ means inhibiting the synthesis (e.g. byinhibiting transcription, or translation) and/or inhibiting thesecretion in a cell that has the ability to produce and/or secrete thesecytokines.

As used herein, a composition that “substantially” comprises a compoundmeans that the composition contains more than about 80% by weight, morepreferably more than about 90% by weight, even more preferably more thanabout 95% by weight, and most preferably more than about 97% by weightof the compound.

As used herein, a composition that is “substantially free” of a compoundmeans that the composition contains less than about 20% by weight, morepreferably less than about 10% by weight, even more preferably less thanabout 5% by weight, and most preferably less than about 3% by weight ofthe compound.

As used herein, a reaction that is “substantially complete” means thatthe reaction contains more than about 80% by weight of the desiredproduct, more preferably more than about 90% by weight of the desiredproduct, even more preferably more than about 95% by weight of thedesired product, and most preferably more than about 97% by weight ofthe desired product.

As used herein, a racemic mixture means about 50% of one enantiomer andabout 50% of is corresponding enantiomer relative to all chiral centersin the molecule. The invention encompasses all enantiomerically-pure,enantiomerically-enriched, diastereomerically pure, diastereomericallyenriched, and racemic mixtures of the compounds of Table 1.

Enantiomeric and diastereomeric mixtures can be resolved into theircomponent enantiomers or stereoisomers by well known methods, such aschiral-phase gas chromatography, chiral-phase high performance liquidchromatography, crystallizing the compound as a chiral salt complex, orcrystallizing the compound in a chiral solvent. Enantiomers anddiastereomers can also be obtained from diastereomerically- orenantiomerically-pure intermediates, reagents, and catalysts by wellknown asymmetric synthetic methods.

When administered to a patient, e.g., to a non-human animal forveterinary use or for improvement of livestock, or to a human forclinical use, the compounds of the invention are typically administeredin isolated form or as the isolated form in a pharmaceuticalcomposition. As used herein, “isolated” means that the compounds of theinvention are separated from other components of either (a) a naturalsource, such as a plant or cell, preferably bacterial culture, or (b) asynthetic organic chemical reaction mixture. Preferably, viaconventional techniques, the compounds of the invention are purified. Asused herein, “purified” means that when isolated, the isolate containsat least 95%, preferably at least 98%, of a single compound of theinvention by weight of the isolate.

Only those choices and combinations of substituents that result in astable structure are contemplated. Such choices and combinations will beapparent to those of ordinary skill in the art and may be determinedwithout undue experimentation.

The invention can be understood more fully by reference to the followingdetailed description and illustrative examples, which are intended toexemplify non-limiting embodiments of the invention.

Specific Embodiments

The invention relates to compounds and pharmaceutical compositions thatare particularly useful for immunosuppression or to treat or preventinflammatory conditions, immune disorders, and allergic disorders.

In one embodiment, the invention relates to compounds selected from thegroup consisting of:

-   2,6-difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(thiazol-2-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   2,6-difluoro-N-(4-(3-methyl-5-(pyridin-3-yl)thiophen-2-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(pyridin-3-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1,3,4-oxadiazol-2-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(6-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamide;-   N-(4-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(4-methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamide;-   N-(4-(5-(5-chloro-2-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;-   3-methyl-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamide;-   N-(4-(5-(4-chloropyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(5-(4-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;-   N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide;-   2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide    hydrochloride; or-   N-(4-(5-(1-ethyl-1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;    -   or pharmaceutically acceptable salts, solvates, clathrates, or        prodrugs thereof.

In one aspect, the invention is2,6-difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(3-methyl-5-(thiazol-2-yl)thiophen-2-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(3-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-2-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(3-methyl-5-(pyridin-3-yl)thiophen-2-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(pyridin-3-yl)thiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(1,3,4-oxadiazol-2-yl)thiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(6-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention isN-(4-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(4-methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention isN-(4-(5-(5-chloro-2-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyrazin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention isN-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is3-methyl-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention isN-(4-(5-(4-chloropyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(5-(4-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention isN-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In one aspect, the invention is2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamidehydrochloride or a pharmaceutically acceptable salt, solvate, clathrate,or prodrug thereof.

In one aspect, the invention isN-(4-(5-(1-ethyl-1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamideor a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

All of the features, specific embodiments and particular substituentsdisclosed herein may be combined in any combination. Each feature,embodiment or substituent disclosed in this specification may bereplaced by an alternative feature, embodiment or substituent servingthe same, equivalent, or similar purpose. In the case of chemicalcompounds, specific values for variables (e.g., values shown in theexemplary compounds disclosed herein) in any chemical formula disclosedherein can be combined in any combination resulting in a stablestructure. Furthermore, specific values (whether preferred or not) forsubstituents in one type of chemical structure may be combined withvalues for other substituents (whether preferred or not) in the same ordifferent type of chemical structure. Thus, unless expressly statedotherwise, each feature, embodiment or substituent disclosed is only anexample of a generic series of equivalent or similar features,embodiments or substituents.

In some embodiments, the invention relates to pharmaceuticalcompositions that comprise a compound of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof, as an activeingredient, and a pharmaceutically acceptable carrier or vehicle. Thecompositions are useful for immunosuppression or to treat or preventinflammatory conditions, allergic conditions and immune disorders.

In some embodiments, the invention relates to methods forimmunosuppression or for treating or preventing inflammatory conditions,immune disorders, or allergic disorders in a patient in need thereofcomprising administering an effective amount of a compound of Table 1,or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In some embodiments, the invention relates to methods forimmunosuppression or for treating or preventing inflammatory conditions,immune disorders, or allergic disorders in a patient in need thereofcomprising administering an effective amount of a pharmaceuticalcomposition that comprises a compound of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof.

In some embodiments, compounds of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof, areparticularly useful inhibiting immune cell (e.g., T-cells and/orB-cells) activation (e.g., activation in response to an antigen) and/orT cell and/or B cell proliferation. Indicators of immune cell activationinclude secretion of IL-2 by T cells, proliferation of T cells and/or Bcells, and the like. In one embodiment, immune cell activation and/or Tcell and/or B cell proliferation is inhibited in a mammal (e.g., ahuman), by administering to the mammal (e.g., human) a compound of Table1, or a pharmaceutically acceptable salt, solvate, clathrate, or prodrugthereof.

In some embodiments, compounds of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof, can inhibit theproduction of certain cytokines that regulate immune cell activation.For example, compounds of Table 1, or a pharmaceutically acceptablesalt, solvate, clathrate, or prodrug thereof, can inhibit the productionof IL-2, IL-4, IL-5, IL-13, GM-CSF, IFN-γ, TNF-α and combinationsthereof. In one embodiment, cytokine production is inhibited in a mammal(e.g., a human), by administering to the mammal (e.g., human) a compoundof Table 1, or a pharmaceutically acceptable salt, solvate, clathrate,or prodrug thereof.

In some embodiments, compounds of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof, can modulatethe activity of one or more ion channel, such as CRAC ion channels,involved in activation of immune cells. In some embodiments, a compoundof Table 1 can inhibit the influx of calcium ions into an immune cell(e.g., T cells, B cells, and/or mast cells) by inhibiting the action ofCRAC ion channels. In general, a decrease in I_(CRAC) current uponcontacting a cell with a compound is one indicator that the compoundinhibitions CRAC ion channels. I_(CRAC) current can be measured, forexample, using a patch clamp technique, which is described in moredetail in the examples below. In some embodiments, a compound of Table 1modulates an ion channel in a mammal (e.g., a human). In someembodiments, the activity of one or more ion channels is inhibited in amammal (e.g., a human), by administering to the mammal (e.g., human) acompound of Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof.

In some embodiments, compounds of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof, can inhibitdegranulation of mast cell. Inhibition of mast cell degranulation candetermined as described in the experimental section herein or by anymethod known to those skilled in the art. In some embodiments, mast celldegranulation is inhibited in a mammal (e.g., a human), by administeringto the mammal (e.g., human) a compound of Table 1, or a pharmaceuticallyacceptable salt, solvate, clathrate, or prodrug thereof.

Exemplary Compounds of the Invention

Exemplary compounds of the invention are depicted in Table 1 below.

TABLE 1 No. Structure Name 1

2,6-difluoro-N-(4-(5-(isoxazol-5-yl)- 3-methylthiophen-2-yl)phenyl)benzamide 2

2,6-difluoro-N-(4-(3-methyl-5- (thiazol-2-yl)thiophen-2-yl)phenyl)benzamide 3

2,6-difluoro-N-(4-(3-methyl-5-(1- methyl-1H-imidazol-5-ypthiophen-2-yl)phenyl)benzamide 4

2,6-difluoro-N-(5-(2-methyl-5- (oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide 5

2,6-difluoro-N-(5-(2-methyl-5- (oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide 6

2,6-difluoro-N-(4-(3-methyl-5- (pyridin-3-yl)thiophen-2-yl)phenyl)benzamide 7

2,6-difluoro-N-(4-(2-methyl-5- (pyridin-3-yl)thiophen-3-yl)phenyl)benzamide 8

2,6-difluoro-N-(4-(2-methyl-5- (thiazol-2-ypthiophen-3-yl)phenyl)benzamide 9

2,6-difluoro-N-(4-(2-methyl-5-(1,3,4- oxadiazol-2-yl)thiophen-3-yl)phenyl)benzamide 10

2,6-difluoro-N-(6-(2-methyl-5- (oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamide 11

N-(4-(5-(1H-imidazol-5-yl)-2- methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide 12

2,6-difluoro-N-(4-(2-methyl-5-(4- methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamide 13

N-(4-(5-(5-chloro-2-methoxypyridin- 3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide 14

2,6-difluoro-N-(5-(2-methyl-5- (thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamide 15

2,6-difluoro-N-(5 -(2-methyl-5- (thiazol-2-ypthiophen-3-yl)pyrazin-2-yl)benzamide 16

N-(5-(5-(1H-imidazol-5-yl)-2- methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamide 17

2,6-difluoro-N-(5-(2-methyl-5-(1- methyl-1H-imidazol-5-yl)thiophen-3 -yl)pyridin-2-yl)benzamide 18

3-methyl-N-(5-(2-methyl-5-(oxazol- 5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamide 19

N-(4-(5-(4-chloropyridin-3-yl)-2- methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide 20

2,6-difluoro-N-(4-(5-(4- methoxypyridin-3 -yl)-2- methylthiophen-3-yl)phenyl)benzamide 21

2,6-difluoro-N-(5-(2-methyl-5-(1- methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide 22

N-(5-(5-(1H-imidazol-5-yl)-2- methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamide 23

2,6-difluoro-N-(4-(2-methyl-5-(1- methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide 24

2,6-difluoro-N-(4-(2-methyl-5-(1- methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide hydrochloride 25

N-(4-(5-(1-ethyl-1H-imidazol-5-yl)- 2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide

Mechanism of Action

Activation of T-lymphocytes in response to an antigen is dependent oncalcium ion oscillations. Calcium ion oscillations in T-lymphocytes aretriggered through stimulation of the T-cell antigen receptor, andinvolve calcium ion influx through the stored-operatedCa²⁺-release-activated Ca²⁺ (CRAC) channel. In addition, antigen induceddegranulation of mast cells has also been shown to be initiated bycalcium ion in flux. Although the molecular structure of the CRAC ionchannel has not been identified, a detailed electrophysiological profileof the channel exist. Thus, inhibition of CRAC ion channels can bemeasured by measuring inhibition of the I_(CRAC) current. Calcium ionoscillations in T-cells have been implicated in the activation ofseveral transcription factors (e.g., NFAT, Oct/Oap and NFκB) which arecritical for T-cell activation (Lewis, Biochemical Society Transactions(2003), 31:925-929, the entire teachings of which are incorporatedherein by reference). Without wishing to be bound by any theory, it isbelieved that because the compounds of the invention inhibit theactivity of CRAC ion channels, they inhibit immune cell activation.

Methods of Treatment and Prevention

In accordance with the invention, an effective amount of a compound ofTable 1, or a pharmaceutically acceptable salt, solvate, clathrate, andprodrug thereof, or a pharmaceutical composition comprising a compoundof Table 1, or a pharmaceutically acceptable salt, solvate, clathrate,and prodrug thereof, is administered to a patient in need ofimmunosuppression or in need of treatment or prevention of aninflammatory condition, an immune disorder, or an allergic disorder.Such patients may be treatment naïve or may experience partial or noresponse to conventional therapies.

Responsiveness to immunosuppression or of a particular inflammatorycondition, immune disorder, or allergic disorder in a subject can bemeasured directly (e.g., measuring blood levels of inflammatorycytokines (such as IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α, IFN-γ and thelike) after administration of a compound of this invention), or can beinferred based on an understanding of disease etiology and progression.The compounds of Table 1, or pharmaceutically acceptable salts,solvates, clathrates, and prodrugs thereof can be assayed in vitro or invivo, for the desired therapeutic or prophylactic activity, prior to usein humans. For example, known animal models of inflammatory conditions,immune disorders, or allergic disorders can be used to demonstrate thesafety and efficacy of compounds of this invention.

Preparation of Compounds of the Invention

In general, the phenyl and pyridinyl compounds of the invention thathave amide linkers are prepared by contacting a[1,3,2]dioxaborolan-2-yl-phenyl or -pyridinyl derivative (XVIII) with anacid chloride (XVI) in the presence of a base to form intermediatecompound (VI) having an amide linkage (see Scheme I). Typically, anaprotic solvent and aprotic base is used in this reaction.

Intermediate (VI) is then reacted with a halo-heteroaryl derivative(VII) in the presence of a palladium catalyst and a base (Suzukicoupling reaction) to form a phenyl or pyridinyl compound of theinvention having amide linkers (VIII) (see Scheme II).

Phenyl or pyridinyl compounds of the invention having an amide linker inwhich the amine group is attached to Y and the carbonyl group isattached to the Phenyl or pyridinyl ring can be prepared by reacting4-halo-benzoyl chloride or a 5-halo-pyridine-2-carbonyl chloride (IX)with an amine derivative (X) in the presence of a base to formintermediate compound (XI) (see Scheme III).

Intermediate (XI) is then reacted with a boric acid derivative (XII) inthe presence of PdCl₂(PPh₃)₄ and a base (Suzuki Coupling reaction, as inScheme IV) to form phenyl or pyridinyl compound of the invention (XIII).

Compounds of the invention in which L is —NHC(S)— or —C(S)NH— can beprepared by treating compounds having an amide linker with Lawesson'sreagent.

Compounds of the invention having —CH₂—NH— or —NH—CH₂— linkers can beprepared by contacting compounds having —NHC(S)— or —C(S)NH— linkerswith Raney Ni. Alternatively, compounds of the invention having a—CH₂—NH— or —NH—CH₂— linker can be prepared by reducing a compoundhaving a —C(O)—NH— or —NH—C(O)— linker, respectively, with, for example,sodium borohydride. Alternatively, compounds that have —NHCH₂— linkerscan be prepared by reacting aldehyde (f) with amine (XX) followed byreduction of the shift base with sodium borohydride as shown in SchemeIVa (see U.S. patent application Ser. No. 10/897,681, filed on Jul. 22,2004, the entire teachings of which are incorporated herein byreference).

Compounds of the invention having —C(O)— linkers can be prepared by aFriedel-Craft acylation reaction by reacting a halo-phenyl orhalo-pyridinyl derivative (XIV) with an acid chloride (XV) in thepresence of AlCl₃ to form an intermediate which can then be reacted withan [1,3,2]dioxaborolan-2-yl-heteroaryl (XVI) in the presence of apalladium catalyst and a base to form a compound of the invention havinga carbonyl linker (XVII) (see Scheme V).

Compounds of the invention that have —C(S)— can be prepared fromcompounds that have carbonyl linkers by treating them with Lawesson'sreagent or P₂S₅ in pyridine.

Compounds of the invention that have a sulfonamide linker (XXII) can beprepared by reacting an amine derivative (XX), which is prepared by ananalogous method as described in Scheme II, with a sulfonyl chloridederivative (XXI) as shown in Scheme VI. Typically, the amine derivative(XX) is dissolved in a polar solvent, such as an alcohol, and thesulfonyl chloride derivative (XXI) is added. The reaction is typicallyheated to about 50° C. to about 100° C.

Compounds of the invention having a urea linker (XXIV) can be preparedby reacting amine derivative (XX) with an isocyanate (XXIII) as shown inScheme VII. Typically, the amine derivative (XX) is dissolved in anon-polar, aprotic solvent such as dichloromethane (DCM) to which theisocyanate (XXIII) is added at room temperature. The reaction istypically stirred for about 5 minutes to about 1 hour to give a compoundof the invention having a urea linker (XXIV)

Compounds of the invention having a thiourea linker (—NHC(S)NH—) can beprepared by treating compounds having a urea linker with Lawesson'sreagent.

Compounds of the invention having a hydrazinyl linker (—NH—N═CH—) can beprepared by adding an aqueous solution of NaNO₂ (1 eq.) to a solution ofamine derivative (XX) (1 eq.) in concentrated HCl at about 0° C. Afterthe solution is stirred at about 0° C. for about 15 minute to about 1hour, then 2.4 eq. of SnCl₂ in concentrated HCl is added, and thereaction is stirred at about 0° C. for about 1 hour to give ahydrazinium chloride intermediate (XXV). The hydrazinium chlorideintermediate (XXV) is dissolved in acetic acid and an alcohol, such asmethanol, and an aldehyde (XXVI) is added. The reaction is stirred atroom temperature for about an hour to give a compound of the inventionhaving a hydrazinyl linker (XXVII) (see Scheme VIII).

Compounds of the invention that have a double bond linker can beprepared by heating a mixture of a 4-halo-benzyl halide or ahalomethyl-halo-pyridine (XXVIII) and a trialkyl-phosphite, such astriethyl phosphate, in a non-polar, aprotic solvent to form a dialkylphosphate derivative (XXIX). The dialkyl phosphate derivative (XXIX) isthen dissolved in a polar, aprotic solvent, such as an ether, and cooledto about −25° C. to about −78° C. and sodium-hexamethyldisilazane(NaHMDS) is added. After about 5 minutes to about 30 minutes an aldehydeis added and the solution is stirred for about 15 minutes to about 1hour then allowed to warm to room temperature. The reaction is quenchedwith an aqueous ammonium chloride solution to form alkene intermediate(XXX). Alkene intermediate (XXX) is then coupled with cycloalkyleneboronic acid ester (XVII) in a similar manner as described in Scheme IIto form a compound of the invention that has a double bond linker (XXXI)(see Scheme IX).

Compounds that have an amine linker (XXXII) can be prepared by stirringa mixture of amine derivative (XX) (1 equ.), triphenylbismuthine(III)(1.1-1.5 equ.) and Cu(OAc)₂ (1.1-1.5 equ.) in dichloromethane at roomtemperature for about 2-12 hours (see Scheme X).

Pharmaceutical Compositions and Dosage Forms

Pharmaceutical compositions and dosage forms of the invention compriseone or more active ingredients in relative amounts and formulated insuch a way that a given pharmaceutical composition or dosage form can beused for immunosuppression or to treat or prevent inflammatoryconditions, immune disorders, and allergic disorders. Preferredpharmaceutical compositions and dosage forms comprise a compound ofTable 1, or a pharmaceutically acceptable prodrug, salt, solvate, orclathrate thereof, optionally in combination with one or more additionalactive agents.

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), or transdermal administration to a patient. Examples ofdosage forms include, but are not limited to: tablets; caplets;capsules, such as soft elastic gelatin capsules; cachets; troches;lozenges; dispersions; suppositories; ointments; cataplasms (poultices);pastes; powders; dressings; creams; plasters; solutions; patches;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage formsuitable for mucosal administration may contain a smaller amount ofactive ingredient(s) than an oral dosage form used to treat the sameindication. This aspect of the invention will be readily apparent tothose skilled in the art. See, e.g., Remington's Pharmaceutical Sciences(1990) 18th ed., Mack Publishing, Easton Pa.

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms.

The suitability of a particular excipient may also depend on thespecific active ingredients in the dosage form. For example, thedecomposition of some active ingredients can be accelerated by someexcipients such as lactose, or when exposed to water. Active ingredientsthat comprise primary or secondary amines (e.g., N-desmethylvenlafaxineand N,N-didesmethylvenlafaxine) are particularly susceptible to suchaccelerated decomposition. Consequently, this invention encompassespharmaceutical compositions and dosage forms that contain little, ifany, lactose. As used herein, the term “lactose-free” means that theamount of lactose present, if any, is insufficient to substantiallyincrease the degradation rate of an active ingredient. Lactose-freecompositions of the invention can comprise excipients that are wellknown in the art and are listed, for example, in the U.S. Pharmocopia(USP)SP (XXI)/NF (XVI). In general, lactose-free compositions compriseactive ingredients, a binder/filler, and a lubricant in pharmaceuticallycompatible and pharmaceutically acceptable amounts. Preferredlactose-free dosage forms comprise active ingredients, microcrystallinecellulose, pre-gelatinized starch, and magnesium stearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen (1995) Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 379-80. In effect, water andheat accelerate the decomposition of some compounds. Thus, the effect ofwater on a formulation can be of great significance since moistureand/or humidity are commonly encountered during manufacture, handling,packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizer” include, but are not limited to, antioxidantssuch as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms of the invention comprise acompound of Table 1, or a pharmaceutically acceptable salt, solvate,clathrate, or prodrug thereof in an amount of from about 1 mg to about1000 mg, preferably in an amount of from about 50 mg to about 500 mg,and most preferably in an amount of from about 75 mg to about 350 mg.The typical total daily dosage of a compound of Table 1, or apharmaceutically acceptable salt, solvate, clathrate, or prodrug thereofcan range from about 0.001 mg to about 5000 mg per day, preferably in anamount from about 0.01 mg to about 1500 mg per day, more preferably fromabout 0.01 mg to about 1000 mg per day. It is within the skill of theart to determine the appropriate dose and dosage form for a givenpatient.

Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally, Remington's Pharmaceutical Sciences (1990) 18th ed., MackPublishing, Easton Pa.

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an admixture with at least one excipientaccording to conventional pharmaceutical compounding techniques.Excipients can take a wide variety of forms depending on the form ofpreparation desired for administration. For example, excipients suitablefor use in oral liquid or aerosol dosage forms include, but are notlimited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101, AVICEL-PH-103 AVICELRC-581, AVICEL-PH-105 (available from FMC Corporation, American ViscoseDivision, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof. Onespecific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581. Suitable anhydrous or lowmoisture excipients or additives include AVICEL-PH-103J and Starch 1500LM.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,preferably from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, other starches, pre-gelatinizedstarch, other starches, clays, other algins, other celluloses, gums, andmixtures thereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about 1 weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

Controlled Release Dosage Forms

Active ingredients of the invention can be administered by controlledrelease means or by delivery devices that are well known to those ofordinary skill in the art. Examples include, but are not limited to,those described in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809;3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767, 5,120,548,5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of which isincorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

A particular extended release formulation of this invention comprises atherapeutically or prophylactically effective amount of a compound ofTable 1, or a pharmaceutically acceptable salt, solvate, hydrate,clathrate, or prodrug thereof, in spheroids which further comprisemicrocrystalline cellulose and, optionally,hydroxypropylmethyl-cellulose coated with a mixture of ethyl celluloseand hydroxypropylmethylcellulose. Such extended release formulations canbe prepared according to U.S. Pat. No. 6,274,171, the entire teachingsof which are incorporated herein by reference.

A specific controlled-release formulation of this invention comprisesfrom about 6% to about 40% a compound of Table 1 by weight, about 50% toabout 94% microcrystalline cellulose, NF, by weight, and optionally fromabout 0.25% to about 1% by weight of hydroxypropyl-methylcellulose, USP,wherein the spheroids are coated with a film coating compositioncomprised of ethyl cellulose and hydroxypropylmethylcellulose.

Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

Transdermal, Topical, and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms of the invention include,but are not limited to, ophthalmic solutions, sprays, aerosols, creams,lotions, ointments, gels, solutions, emulsions, suspensions, or otherforms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa. and Introduction to Pharmaceutical Dosage Forms(1985) 4th ed., Lea & Febiger, Philadelphia. Dosage forms suitable fortreating mucosal tissues within the oral cavity can be formulated asmouthwashes or as oral gels. Further, transdermal dosage forms include“reservoir type” or “matrix type” patches, which can be applied to theskin and worn for a specific period of time to permit the penetration ofa desired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed by this invention are well known to those skilled inthe pharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof to form lotions, tinctures, creams, emulsions, gelsor ointments, which are non-toxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences (1980 & 1990) 16th and 18th eds., MackPublishing, Easton Pa.

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, oleyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80(polysorbate 80) and Span 60 (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different salts, hydrates or solvates ofthe active ingredients can be used to further adjust the properties ofthe resulting composition.

Combination Therapy

The methods for immunosuppression or for treating or preventinginflammatory conditions, allergic disorders, and immune disorders in apatient in need thereof can further comprise administering to thepatient being administered a compound of this invention, an effectiveamount of one or more other active agents. Such active agents mayinclude those used conventionally for immunosuppression or forinflammatory conditions, allergic disorders, or immune disorders. Theseother active agents may also be those that provide other benefits whenadministered in combination with the compounds of this invention. Forexample, other therapeutic agents may include, without limitation,steroids, non-steroidal anti-inflammatory agents, antihistamines,analgesics, immunosuppressive agents and suitable mixtures thereof. Insuch combination therapy treatment, both the compounds of this inventionand the other drug agent(s) are administered to a subject (e.g., humans,male or female) by conventional methods. The agents may be administeredin a single dosage form or in separate dosage forms. Effective amountsof the other therapeutic agents and dosage forms are well known to thoseskilled in the art. It is well within the skilled artisan's purview todetermine the other therapeutic agent's optimal effective-amount range.

In one embodiment of the invention where another therapeutic agent isadministered to a subject, the effective amount of the compound of thisinvention is less than its effective amount when the other therapeuticagent is not administered. In another embodiment, the effective amountof the conventional agent is less than its effective amount when thecompound of this invention is not administered. In this way, undesiredside effects associated with high doses of either agent may beminimized. Other potential advantages (including without limitationimproved dosing regimens and/or reduced drug cost) will be apparent tothose of skill in the art.

In one embodiment relating to autoimmune, allergic and inflammatoryconditions, the other therapeutic agent may be a steroid or anon-steroidal anti-inflammatory agent. Particularly useful non-steroidalanti-inflammatory agents, include, but are not limited to, aspirin,ibuprofen, diclofenac, naproxen, benoxaprofen, flurbiprofen, fenoprofen,flubufen, ketoprofen, indoprofen, piroprofen, carprofen, oxaprozin,pramoprofen, muroprofen, trioxaprofen, suprofen, aminoprofen,tiaprofenic acid, fluprofen, bucloxic acid, indomethacin, sulindac,tolmetin, zomepirac, tiopinac, zidometacin, acemetacin, fentiazac,clidanac, oxpinac, mefenamic acid, meclofenamic acid, flufenamic acid,niflumic acid, tolfenamic acid, diflurisal, flufenisal, piroxicam,sudoxicam, isoxicam; salicylic acid derivatives, including aspirin,sodium salicylate, choline magnesium trisalicylate, salsalate,diflunisal, salicylsalicylic acid, sulfasalazine, and olsalazin;para-aminophennol derivatives including acetaminophen and phenacetin;indole and indene acetic acids, including indomethacin, sulindac, andetodolac; heteroaryl acetic acids, including tolmetin, diclofenac, andketorolac; anthranilic acids (fenamates), including mefenamic acid, andmeclofenamic acid; enolic acids, including oxicams (piroxicam,tenoxicam), and pyrazolidinediones (phenylbutazone, oxyphenthartazone);and alkanones, including nabumetone and pharmaceutically acceptablesalts thereof and mixtures thereof. For a more detailed description ofthe NSAIDs, see Paul A. Insel, Analgesic-Antipyretic andAntiinflammatory Agents and Drugs Employed in the Treatment of Gout, inGoodman & Gilman's The Pharmacological Basis of Therapeutics 617-57(Perry B. Molinhoff and Raymond W. Ruddon eds., 9^(th) ed 1996) and GlenR. Hanson, Analgesic, Antipyretic and Anti-Inflammatory Drugs inRemington: The Science and Practice of Pharmacy Vol II 1196-1221 (A. R.Gennaro ed. 19th ed. 1995) which are hereby incorporated by reference intheir entireties.

Of particular relevance to allergic disorders, the other therapeuticagent may be an antihistamine. Useful antihistamines include, but arenot limited to, loratadine, cetirizine, fexofenadine, desloratadine,diphenhydramine, chlorpheniramine, chlorcyclizine, pyrilamine,promethazine, terfenadine, doxepin, carbinoxamine, clemastine,tripelennamine, brompheniramine, hydroxyzine, cyclizine, meclizine,cyproheptadine, phenindamine, acrivastine, azelastine, levocabastine,and mixtures thereof. For a more detailed description of antihistamines,see Goodman & Gilman's The Pharmacological Basis of Therapeutics (2001)651-57, 10^(th) ed).

Immunosuppressive agents include glucocorticoids, corticosteroids (suchas Prednisone or Solumedrol), T cell blockers (such as cyclosporin A andFK506), purine analogs (such as azathioprine (Imuran)), pyrimidineanalogs (such as cytosine arabinoside), alkylating agents (such asnitrogen mustard, phenylalanine mustard, buslfan, and cyclophosphamide),folic acid antagonsists (such as aminopterin and methotrexate),antibiotics (such as rapamycin, actinomycin D, mitomycin C, puramycin,and chloramphenicol), human IgG, antilymphocyte globulin (ALG), andantibodies (such as anti-CD3 (OKT3), anti-CD4 (OKT4), anti-CD5,anti-CD7, anti-IL-2 receptor, anti-alpha/beta TCR, anti-ICAM-1,anti-CD20 (Rituxan), anti-IL-12 and antibodies to immunotoxins).

The foregoing and other useful combination therapies will be understoodand appreciated by those of skill in the art. Potential advantages ofsuch combination therapies include a different efficacy profile, theability to use less of each of the individual active ingredients tominimize toxic side effects, synergistic improvements in efficacy,improved ease of administration or use and/or reduced overall expense ofcompound preparation or formulation.

Other Embodiments

The compounds of this invention may be used as research tools (forexample, as a positive control for evaluating other potential CRACinhibitors, or IL-2, IL-4, IL-5, IL-13, GM-CSF, TNF-α, and/or INF-γinhibitors). These and other uses and embodiments of the compounds andcompositions of this invention will be apparent to those of ordinaryskill in the art.

The invention is further defined by reference to the following examplesdescribing in detail the preparation of compounds of the invention. Itwill be apparent to those skilled in the art that many modifications,both to materials and methods, may be practiced without departing fromthe purpose and interest of this invention. The following examples areset forth to assist in understanding the invention and should not beconstrued as specifically limiting the invention described and claimedherein. Such variations of the invention, including the substitution ofall equivalents now known or later developed, which would be within thepurview of those skilled in the art, and changes in formulation or minorchanges in experimental design, are to be considered to fall within thescope of the invention incorporated herein.

EXAMPLES Experimental Rationale

Without wishing to be bound by theory, it is believed that the compoundsof this invention inhibit CRAC ion channels, thereby inhibitingproduction of IL-2 and other key cytokines involved with inflammatory,allergic and immune responses. The examples that follow demonstratethese properties.

Materials and General Methods

Reagents and solvents used below can be obtained from commercial sourcessuch as Aldrich Chemical Co. (Milwaukee, Wis., USA). ¹H-NMR and ¹³C-NMRspectra were recorded on a Varian 300 MHz NMR spectrometer. Significantpeaks are tabulated in the order: δ (ppm): chemical shift, multiplicity(s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br s,broad singlet), coupling constant(s) in Hertz (Hz) and number ofprotons.

Patch clamp experiments were performed in the tight-seal whole-cellconfiguration at 21-25° C. High resolution current recordings wereacquired by a computer-based patch clamp amplifier system (EPC-9, HEKA,Lambrecht, Germany). Patch pipettes had resistances between 2-4 MΩ afterfilling with the standard intracellular solution. Immediately followingestablishment of the whole-cell configuration, voltage ramps of 50-200ms duration spanning the voltage range of −100 to +100 mV were deliveredat a rate of 0.5 Hz over a period of 300-400 seconds. All voltages werecorrected for a liquid junction potential of 10 mV between external andinternal solutions when using glutamate as the intracellular anion.Currents were filtered at 2.9 kHz and digitized at 10 μs intervals.Capacitive currents and series resistance were determined and correctedbefore each voltage ramp using the automatic capacitance compensation ofthe EPC-9. The low resolution temporal development of membrane currentswas assessed by extracting the current amplitude at −80 mV or +80 mVfrom individual ramp current records.

Synthesis of Representative Exemplary Compounds of this Invention

In general, the compounds of the invention can be synthesized usingmethods analogous to those described in U.S. patent application Ser. No.10/897,681 and U.S. patent application Ser. No. 11/233,224, the entireteachings of these patent applications are incorporated herein byreference.

Example 14-[4-(2,6-Difluoro-benzoylamino)-phenyl]-5-methyl-thiophene-2-carboxylicacid methyl ester

Step A:

To a stirred solution of4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenylamine (a) (5.2 g,24 mmol), TEA (5 mL) in dry DCM (50 mL) at 0° C. was added2,6-difluoro-benzoyl chloride (b) (3.0 mL, 24 mmol) dropwise. Themixture was allowed to warm to room temperature over 2 h before it waswashed with water (2×100 mL) and dried. Removal of solvents gave2,6-difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamide(c) (8.4 g, 23 mmol) as white solid.

¹H-NMR (CDCl₃) δ (ppm) 7.8 (d, 2H, J=8), 7.7 (br, 1H), 7.6 (m, 2H), 7.4(m, 1H), 7.0 (t, 2H, J=9), 1.35 (s, 12H); ESMS clcd for C₁₉H₂₀BF₂NO₃:359.1. Found: 360.1 (M+H)⁺.

Step B:

A mixture of 4-Iodo-5-methyl-thiophene-2-carboxylic acid methyl ester(d, 1 mmol),2,6-Difluoro-N-[4-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenyl]-benzamide(c, 1 mmol), palladium catalyst (0.1 mmol), sodium bicarbonate (1 mmol)in a mixture of toluene (5 mL), water (1 mL), ethanol (1 mL) was heatedat 100° C. for 24 h. The mixture was taken up with EtOAc (100 mL),washed with water (2×100 mL) and dried (Na₂SO₄). The oil obtained onconcentration was purified by flash chromatography to give4-[4-(2,6-Difluoro-benzoylamino)-phenyl]-5-methyl-thiophene-2-carboxylicacid methyl ester as a yellowish solid (50 mg).

¹H-NMR (CDCl₃) δ 7.8 (br, 1H), 7.7 (m, 3H), 7.4 (m, 3H), 7.0 (t, 2H,J=8) 3.88 (s, 3H), 2.54 (s, 3H) ppm; ESMS calcd for C₂₀H₁₅F₂NO₃S: 387.1.Found: 388.1 (M+H⁺).

Example 23-Methyl-N-[4-(3-methyl-5-pyrimidin-5-yl-thiophen-2-yl)-phenyl]-isonicotinamide

The pyrimidine substituent on the thiophene ring of3-Methyl-N-[4-(3-methyl-5-pyrimidin-5-yl-thiophen-2-yl)-phenyl]-isonicotinamidewas attached using a Suzuki coupling reaction (as describe in Step B ofthe synthesis of compound 1) by reacting a boric acid derivative ofthiophene (o) with 5-bromo-pyridine (p) in the presence of a palladiumcatalyst to form 2-(pyrimidin-5-yl)-4-methyl-thiophene (q). In general,aromatic substituents such as pyridine, can be added to thiophene,oxazole, thiazole and oxazole ring systems by using a Suzuki couplingreaction. A bromo substituent was added to (q) by reacting it withN-bromo-succinimide in acetic acid to form2-(pyrimidin-5-yl)-4-methyl-5-bromo-thiophene (r). Compound (r) is thencoupled to an amino pyridine using a Suzuki coupling reaction (asdescribe in Step B of the synthesis of compound 1) to form Compound (s).Compound (s) is then reacted with 2-methyl-isonicotinoyl chloride in areaction analogous to the reaction described in step A of the synthesisof4-[4-(2,6-Difluoro-benzoylamino)-phenyl]-5-methyl-thiophene-2-carboxylicacid methyl ester to form3-Methyl-N-[4-(3-methyl-5-pyrimidin-5-yl-thiophen-2-yl)-phenyl]-isonicotinamide.

¹H NMR (300 MHz, CDCl₃) δ 9.10 (s, 1H), 8.93 (s, 2H), 8.57-8.54 (m, 2H),7.80 (s, 1H), 7.73 (d, J=8.7 Hz, 2H), 7.53 (d, J=8.7 Hz, 2H), 7.36 (d,J=5.1 Hz, 1H), 7.27 (s, 1H), 2.51 (s, 3H), 2.38 (s, 3H); ESMS cacld(C₂₂H₁₈N₄OS): 386.1. Found: 387.2 (M+H).

Example 3 Methyl4-(4-(2,6-difluorobenzamido)phenyl)-5-methylfuran-2-carboxylate

To a solution of 2,3-dibromo-furan-5-carboxylic acid methyl ester (200mg, 0.70 mmol) in THF (4 mL) was added Pd(PPh₃)₂Cl₂ (50 mg) and MeZnCl(2 M in THF, 420 μL, 0.84 mmol) at room temperature. The reaction wasstirred at this temperature for 12 hr before the solvent was removed.Column chromatography afforded 4-Bromo-5-methyl-furan-2-carboxylic acidmethyl ester (130 mg, 84%). Suzuki coupling of4-Bromo-5-methyl-furan-2-carboxylic acid methyl ester with thecorresponding boronic acid (See the synthesis of Example 1, Step B)provided Methyl4-(4-(2,6-difluorobenzamido)phenyl)-5-methylfuran-2-carboxylate.

¹H NMR (300 MHz, CDCl₃) δ 7.78-7.31 (m, 7H), 7.03-6.96 (m, 2H), 3.91 (s,3H), 2.52 (s, 3H); ESMS cacld (C₂₀H₁₅F₂NO₄): 371.1. Found: 372.2 (M+H).

Example 41-(2,6-difluoro-phenyl)-3-[4-(5-isoxazol-5-yl-3-methyl-thiophen-2-yl)-phenyl]-urea

To 70 mL dichloromethane was added AlCl₃ (2.26 g, 16.9 mmol). Acetylchloride (1.3 mL, 18.2 mmol) was then added drop wise to the abovemixture at 0° C. After stirring at this temperature for 30 min,2-bromo-3-methylthiophene (g) (1 g, 5.65 mmol) was added drop wisethrough a syringe. After stirring at 0° C. for 1 hr, the reaction wasquenched with H₂O (20 mL). The organic layer was washed with NH₄Cl,dried, and concentrated. Purification by silica gel columnchromatography afforded compound (h) in 75% yield.

A solution of (h) (1.33 g, 5.11 mmol) in DMF dimethyl acetal (10 mL) wasrefluxed at 90° C. for 4 hr. The solvent was removed and the residue andhydroxylamine hydrochloride (710 mg, 10.22 mmol) was dissolved inethanol (10 mL). The solution was refluxed at 90° C. for 2 hr. Afterremoval of the volatile components the crude material was purified bysilica gel column chromatography to provide compound (i) in 62% overallyield. Compound (e) was then obtained from (i) and (j) using thestandard Suzuki coupling procedure described above.

To a solution of (e) (40 mg, 0.16 mmol) in dichloromethane (DCM) (2 mL)was added 2,6-difluorophenyl isocyanate (27 mg, 0.18 mmol). The reactionwas stirred at room temperature overnight before it was filtered. Thewhite solid was washed with DCM and methanol to afforded pure1-(2,6-difluoro-phenyl)-3-[4-(5-isoxazol-5-yl-3-methyl-thiophen-2-yl)-phenyl]-urea(35 mg, 53%).

¹H NMR (300 MHz, (CD₃)₂SO) δ 9.16 (s, 1H), 8.62 (s, 1H), 8.18 (s, 1H),7.58-7.43 (m, 3H), 7.46-7.28 (m, 3H), 7.15 (t, J=8.4 Hz, 2H), 6.82 (s,1H), 2.30 (s, 3H); ESMS cacld (C₂₁H₁₅F₂N₃O₂S): 411.1. Found: 412.1(M+H).

Example 51-(2,6-difluoro-phenyl)-3-[4-(5-oxazol-5-yl-3-methyl-thiophen-2-yl)-phenyl]-urea

The mixed solution of 5-bromo-4-methyl-thiophene-2-carbaldehyde (k)(0.93 mmol) in methanol (4 mL) was added tosylmethyl isocyanide (1.02mmol) and K₂CO₃ (1.88 mmol). The reaction was stirred at roomtemperature for 5 min before heated to 80° C. in the sealed tube. After30 min, the solution was cooled to room temperature and concentrated.Column chromatography afforded5-bromo-4-methyl-5-(oxazol-5-yl)-thiophene (l) (190 mg, 80%). Compound(n) was prepared using a Suzuki Coupling reaction analogous to step B ofthe preparation of Example 1.1-(2,6-difluoro-phenyl)-3-[4-(5-oxazol-5-yl-3-methyl-thiophen-2-yl)-phenyl]-ureawas prepared by reacting (n) with 2,6-difluorophenyl isocyanate by ananalogous procedure as that described for Example 4.

¹H NMR (300 MHz, (CD₃)₂SO) δ 9.14 (s, 1H), 8.39 (s, 1H), 8.17 (s, 1H),7.56-7.27 (m, 7H), 7.15 (d, J=8.4 Hz, 2H), 2.28 (s, 3H); ESMS cacld(C₂₁H₁₅F₂N₃O₂S): 411.1. Found: 412.1 (M+H).

Example 62,6-Difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamide

To 70 mL dichloromethane was added AlCl₃ (2.26 g, 16.9 mmol) which waspartially dissolved. Acetyl chloride (1.3 mL, 18.2 mmol) was added dropwise to the above solution at 0° C. After stirring 30 min,2-bromo-3-methylthiophene (1 g, 5.65 mmol) was added drop wise bysyringe. After stirring 1 hr at 0° C., the reaction was quenched withH₂O (20 mL). The organic layer was washed with NH₄Cl, dried, andconcentrated. Column chromatography afforded compound 1 in 75% yield.

The solution of 1 (1.33 g, 5.11 mmol) in DMF dimethyl acetal (10 mL) wasrefluxed at 90° C. for 4 hr. The solvent was removed and the residue andhydroxylamine hydrochloride (710 mg, 10.22 mmol) was dissolved inethanol (10 mL). The solution was refluxed at 90° C. for 2 hr before itwas concentrated. Column chromatography afforded compound 2 in 62%overall yield.2,6-Difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamidewas obtained from 2 following standard procedure; ¹H NMR (300 MHz,CD₃OD) δ 8.39 (d, J=2.1 Hz, 1H), 7.80 (d, J=9.0 Hz, 2H), 7.56-7.46 (m,4H), 7.12 (t, J=8.1 Hz, 2H), 6.61 (d, J=2.1 Hz, 1H), 2.36 (s, 3H); ESMScacld (C₂₁H₁₄F₂N₂O₂S): 396.1. Found: 397.2 (M+H).

Example 72,6-Difluoro-N-(4-(3-methyl-5-(thiazol-2-yl)thiophen-2-yl)phenyl)benzamide

¹H NMR (300 MHz, CDCl₃) δ 8.20 (s, 1H), 7.73-7.70 (m, 3H), 7.49-7.46 (m,2H), 7.42-7.33 (m, 2H), 7.26-7.23 (m, 1H), 6.95 (t, J=8.1 Hz, 2H), 2.33(s, 3H); ESMS cacld (C₂₁H₁₄F₂N₂OS₂): 412.1. Found: 413.1 (M+H).

Example 82,6-Difluoro-N-(4-(3-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-2-yl)phenyl)benzamide

¹H NMR (300 MHz, CD₃OD) δ 7.79-7.75 (m, 2H), 7.70 (s, 1H), 7.59-7.49 (m,3H), 7.15-7.08 (m, 4H), 3.81 (s, 3H), 2.36 (s, 3H); ESMS cacld(C₂₂H₁₇F₂N₃OS): 409.1. Found: 410.2 (M+H).

Example 92,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide(D)

To a solution of 4-bromo-5-methylthiophene-2-carbaldehyde (A, 10.5 mmol)and p-tosylmethyl isocyanide (11 mmol) in methanol (80 mL) was addedpotassium carbonate (13 mmol) and the mixture was heated to 80° C. for 1h. The solvent was removed in vacuo and diluted in DCM (100 mL) andwater (100 mL). The organic layer dried, concentrated and purified byflash chromatography to give 5-(4-bromo-5-methylthiophen-2-yl)oxazole(B, 1.83 g) as yellowish solids.

The above solids (80 mg) and 5-(tributylstannyl)pyrazin-2-amine (97 mg)was dissolved in THF (5 mL), tetrakis(triphenylphosphino)palladium(0) (5mg) was added and the mixture was heated in microwave reactor at 160° C.for 1 h. The solvent was removed and residue was purified by columnchromatography to give5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-amine (C, 16 mg) aswhite solids. This solids was treated with 1 eq. of 2,6-difluorobenzoylchloride in DCM (5 mL) and pyridine (0.1 mL) at rt for 3 h. Removal ofsolvents and purification of residue by flash chromatography gave2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide(D, 12 mg) as white solids. ¹H-NMR (CD₃OD) δ 9.5 (br, 1H), 8.65 (s, 1H),8.18 (s, 1H), 7.70 (s, 1H), 7.5 (m, 2H), 7.32 (s, 1H), 7.1 (t, 2H, J=8),2.75 (s, 3H) ppm; ESMS calcd for C₁₉H₁₂F₂N₄O₂S: 398.1. Found: 399.1(M+H⁺).

Example 102,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide

¹H-NMR (CD₃OD) δ 8.42 (s, 1H), 8.3 (m, 1H), 8.19 (s, 1H), 7.9 (m, 1H),7.5 (m, 1H), 7.42 (s, 1H), 7.32 (s, 1H), 7.1 (t, 2H, J=8), 2.58 (t, 2H,J=8) ppm; ESMS calcd for C₂₀H₁₃F₂N₃O₂S: 397.1. Found: 398.1 (M+H⁺).

Example 112,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide(B)

To a solution of2,6-difluoro-N-(4-(3-methylthiophen-2-yl)phenyl)benzamide (A, 1.3 mmol)in THF (5 mL) was added NBS (1.5 mmol) and solution was stirred at rtfor 2 h. Pyridin-3-ylboronic acid (1.5 mmol),bis(triphenylphosphino)palladium dichloride (0.05 mmol) and NaHCO₃ (3mmol) was added followed by EtOH (2 mL), toluene (5 mL) and water (5mL). The mixture was heated at 110° C. for 1 h. The organic layer wasseparated, concentrated and purified by flash chromatography to give2,6-difluoro-N-(4-(3-methyl-5-(pyridin-3-yl)thiophen-2-yl)phenyl)benzamide(B, 50 mg) as white solids. ¹H-NMR (CD₃OD) □ 8.8 (br, 1H), 8.4 (m, 1H),8.0 (m, 1H), 7.7 (m, 2H), 7.4-7.6 (m, 6H), 7.1 (t, 2H, J=8), 2.38 (s,3H) ppm; ESMS calcd for C₂₃H₁₆F₂N₂OS: 406.1. Found: 407.2 (M+H⁺).

Example 122,6-difluoro-N-(4-(2-methyl-5-(pyridin-3-yl)thiophen-3-yl)phenyl)benzamide

¹H-NMR (CD₃OD) δ 8.8 (br, 1H), 8.4 (m, 2H), 8.1 (m, 1H), 7.8 (m, 2H),7.4-7.7 (m, 4H), 7.2 (m, 1H), 7.1 (t, 2H, J=8), 2.53 (s, 3H) ppm; ESMScalcd for C₂₃H₁₆F₂N₂OS: 406.1. Found: 407.2 (M+H⁺).

Example 132,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide(C)

To a solution of 4-bromo-5-methylthiophene-2-carbonitrile (A, 0.5 g) inMeOH (10 mL) was added 40% aqueous ammonium sulfide solution (1 mL) andthe mixture was stirred at rt for 2 h. The mixture was poured into water(20 mL) and filtered to give 0.67 g intermediate. To the resultingintermediate was added 2-bromo-1,1-diethoxyethane (1 mL) and THF (4 mL)and the mixture was heated in the microwave reactor at 140° C. for 1 h.Removal of solvents and purification by flash chromatography gave2-(4-bromo-5-methylthiophen-2-yl)thiazole as yellow solids (B, 0.46 g).

To the above solids (50 mg) and2,6-difluoro-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)benzamide(50 mg) in a mixture of toluene (2 mL), ethanol (1 mL) and water (2 mL)was added NaHCO₃ (2 mmol) and PdCl₂(PPh₃)₂ (5 mg) and the mixture washeated in microwave reactor at 170° C. for 1 h. The organic layer wasseparated, evaporated and purified by flash chromatography to give2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide(C, 15 mg) as white solids. ¹H-NMR (CD₃OD) □ 7.7 (m, 3H), 7.4-7.6 (m,5H), 7.1 (t, 2H, J=8), 2.57 (s, 3H) ppm; ESMS calcd for C₂₁H₁₄F₂N₂OS₂:412.1. Found: 413.1 (M+H⁺).

Example 142,6-difluoro-N-(4-(2-methyl-5-(1,3,4-oxadiazol-2-yl)thiophen-3-yl)phenyl)-benzamide(C)

To a solution of methyl 4-iodo-5-methylthiophene-2-carboxylate (A, 3.3g) in MeOH (20 mL) was added hydrazine hydrate (0.2 mL) for 12 h. Themixture was cooled to rt and filtered to yield intermediate as whitesolids (1.52 g). The above intermediate (0.5 g) was dissolved in methylorthoformate (5 mL) was heated in microwave reactor at 130° C. for 1 h.The mixture was evaporated and purified by flash chromatography to give2-(4-iodo-5-methylthiophen-2-yl)-1,3,4-oxadiazole (B, 0.10 g) as whitesolids.

To the above solid (B, 30 mg) and2,6-difluoro-N-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)benzamide(30 mg) in a mixture of toluene (2 mL), ethanol (1 mL) and water (2 mL)was added NaHCO₃ (2 mmol) and PdCl₂(PPh₃)₂ (5 mg) and the mixture washeated in microwave reactor at 170° C. for 1 h. The organic layer wasseparated, evaporated and purified by flash chromatography to give2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide(C, 13 mg) as white solids. ¹H-NMR (CD₃OD) □ 8.9 (br, 1H), 7.8 (m, 3H),7.6 (m, 1H), 7.5 (m, 3H), 7.1 (t, 2H, J=8), 2.60 (s, 3H) ppm; ESMS calcdfor C₂₀H₁₃F₂N₃O₂S: 397.1. Found: 398.1 (M+H⁺).

Example 152,6-difluoro-N-(6-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamide

¹H-NMR (CD₃OD) δ 8.8 (br, 1H), 8.3 (m, 1H), 8.18 (s, 1H), 7.5-7.7 (m,4H), 7.32 (s, 1H), 7.1 (t, 2H, J=8), 2.68 (s, 3H) ppm; ESMS calcd forC₂₀H₁₃F₂N₃O₂S: 397.1. Found: 398.1 (M+H⁺).

Example 16N-(4-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide

¹H-NMR (CD₃OD) δ 7.8 (d, 2H, J=8), 7.65 (s, 1H), 7.4-7.6 (m, 4H), 7.3(m, 2H), 7.1 (t, 2H, J=8), 2.48 (s, 3H) ppm; ESMS calcd forC₂₁H₁₅F₂N₃OS: 395.1. Found: 396.1 (M+H⁺).

Example 172,6-difluoro-N-(4-(2-methyl-5-(4-methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamide

¹H-NMR (CD₃OD) δ 8.50 (s, 1H), 8.4 (m, 1H), 7.7 (d, 2H, J=8), 7.5 (m,3H), 7.4 (m, 1H), 7.18 (s, 1H), 7.1 (t, 2H, J=8), 2.55 (s, 3H), 2.52 (s,3H) ppm; ESMS calcd for C₂₄H₁₈F₂N₂OS: 420.1. Found: 421.2 (M+H⁺).

Example 18N-(4-(5-(5-chloro-2-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide

¹H-NMR (CD₃OD) δ 8.0 (m, 1H), 7.8 (m, 2H), 7.4-7.6 (m, 4H), 7.1 (m, 3H),4.08 (s, 3H), 2.54 (s, 3H) ppm; ESMS calcd for C₂₄H₁₇ClF₂N₂O₂S: 470.1.Found: 471.1 (M+H⁺).

Example 192,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamide

¹H-NMR (CD₃OD) δ 8.48 (s, 1H), 8.4 (m, 1H), 8.0 (m, 1H), 7.72 (s, 1H),7.5 (m, 3H), 7.1 (t, 2H, J=8), 2.59 (s, 3H) ppm; ESMS calcd forC₂₀H₁₃F₂N₃OS₂: 413.1. Found: 414.0 (M+H⁺).

Example 202,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyrazin-2-yl)benzamide

¹H-NMR (CD₃OD) δ 9.52 (s, 1H), 8.69 (s, 1H), 7.93 (s, 1H), 7.76 (s, 1H),7.6 (m, 2H), 7.1 (t, 2H, J=8), 2.75 (s, 3H) ppm; ESMS calcd forC₁₉H₁₂F₂N₄OS₂: 414.0. Found: 415.0 (M+H⁺).

Example 21N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamide

ESMS calcd for C₂₀H₁₄F₂N₄OS: 396.1. Found: 397.1 (M+H⁺).

Example 222,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide

¹H-NMR (CD₃OD) δ 8.44 (s, 1H), 7.9 (m, 1H), 7.70 (s, 1H), 7.5 (m, 2H),7.21 (s, 1H), 7.1 (m, 3H), 3.80 (s, 3H), 2.56 (s, 3H) ppm; ESMS calcdfor C₂₁H₁₆F₂N₄OS: 410.1. Found: 411.1 (M+H⁺).

Example 233-methyl-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamide

¹H-NMR (CD₃OD) δ 8.5 (m, 2H), 8.42 (s, 1H), 8.3 (m, 1H), 8.20 (s, 1H),7.9 (m, 1H), 7.5 (m, 1H), 7.46 (s, 1H), 7.32 (s, 1H), 2.55 (s, 3H), 2.18(s, 3H) ppm; ESMS calcd for C₂₀H₁₆N₄O₂S: 376.1. Found: 377.1 (M+H⁺).

Example 24N-(4-(5-(4-chloropyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide

ESMS calcd for C₂₃H₁₅ClF₂N₂OS: 440.1. Found: 441.1 (M+H⁺).

Example 252,6-difluoro-N-(4-(5-(4-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)benzamide

¹H-NMR (CD₃OD) δ, 8.72 (s, 1H), 8.4 (d, 1H, J=5), 7.7 (m, 3H), 7.4 (m,4H), 7.0 (t, 2H, J=8), 6.9 (d, 1H, J=7), 3.99 (s, 3H), 2.55 (s, 3H) ppm;ESMS calcd for C₂₄H₁₈F₂N₂O₂S: 436.1. Found: 437.1 (M+H⁺).

Example 262,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide(B)

To a solution of2,6-difluoro-N-(5-(5-formyl-2-methylthiophen-3-yl)pyrazin-2-yl)benzamide(A, 1.8 mmol) and p-tosylmethyl isocyanide (2 mmol) in methanol (18 mL)was added of a solution of methylamine (4 mmol, 2N in methanol). Themixture was heated in the microwave reactor at 110° C. for 2 h. Removalof solvent and purification of residue by flash chromatography gave2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide(B, 0.24 g) as yellowish solids. ¹H-NMR (CD₃OD) □9.50 (s, 1H), 8.69 (s,1H), 7.70 (s, 1H), 7.5 (m, 2H), 7.1 (m, 3H), 3.71 (s, 3H), 2.72 (s,3H)ppm; ESMS calcd for C₂₀H₁₅F₂N₅OS: 411.1. Found: 412.1 (M+H⁺).

Example 27N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamide

¹H-NMR (CD₃OD) δ, 9.68 (s, 1H), 8.6 (br, 1H), 8.45 (s, 1H), 7.67 (s,1H), 7.50 (s, 1H), 7.5 (m, 1H), 7.23 (s, 1H), 7.0 (t, 2H, J=8), 2.71 (s,3H) ppm; ESMS calcd for C₁₉H₁₃F₂N₅OS: 397.1. Found: 398.1 (M+H⁺).

Example 282,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide(A′) and2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamidehydrochloride (B′)

To the solution of 4-bromo-5-methylthiophene-2-carbaldehyde (1.44 g, 7mmol) in 10 mL of toluene was added 1a′ (2.51 g, 7 mmol), Pd(PPh₃)₂Cl₂(210 mg, 0.3 mmol), NaHCO₃ (1.68 g, 20 mmol), EtOH (2 mL) and water (1mL). The mixture was stirred at 90° C. in a sealed tube under nitrogenatmosphere overnight. The reaction mixture was concentrated and theresidue was partitioned between EtOAc and H₂O. The aqueous phase wasextracted with EtOAc and the combined organic phases were dried(Na₂SO₄), and concentrated. The residue was purified by flashchromatography on silica gel to give 1.50 g of 1a as a yellow solid.ESMS (M+H⁺): 358.

To a solution of 1a (357 mg, 1 mmol) in 5 mL of MeOH was added MeNH₂(2.0 M in MeOH, 2 mL, 4 mmol), TosMIC (450 mg, 1.2 mmol) and silica gel(0.3 g). The mixture was heated to reflux overnight under nitrogenatmosphere. After cooling to the room temperature, the mixture wasconcentrated under reduced pressure, and the residue was purified byflash chromatography on silica gel to give compound A′ (280 mg, 68%) asa yellow solid. A′: ¹H-NMR (CD₃OD) δ 7.67 (d, 2H, J=8.4), 7.42 (s, 1H),7.37-7.27 (m, 3H), 7.03 (s, 1H), 6.95-6.92 (m, 3H), 3.68 (s, 3H), 2.46(s, 3H). ppm; ESMS calcd for C₂₂H₁₇F₂N₃OS: 409.1. Found: 410 (M+H⁺).

To a solution of A′ (100 mg, 0.24 mmol) in 2 mL of EtOAc was added 0.15mL of 2 M HCl in Et₂O. The solution was stirred at room temperature for10 minutes and then stored at 0° C. for 2 hours. The precipitate formedwas collected and dried to give B′ (105 mg) as a white solid. ESMS calcdfor C₂₂H₁₈ClF₂N₃OS: 445. Found: 410 (MH⁺—HCl).

Example 29N-(4-(5-(1-ethyl-1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide

Compound C′ was prepared from compound 1a, ethylamine in ethanolsimilarly as described for the preparation of compound A′.

C′: ¹H-NMR (CDCl₃+CD₃OD) 9.34 (s, 1H, NH), 7.75 (d, 2H, J=8.4),7.47-7.33 (m, 4H), 7.14 (m, 2H), 7.01 (t, 2H, J=8.0), 4.36 (q, 2H, J=7),2.56 (s, 3H), 1.61 (t, 3H, J=7) ppm; ESMS calcd for C₂₃H₁₉F₂N₃OS: 423.1.Found: 424 (M+H⁺).

Example A Inhibition of IL-2 Production

Jurkat cells were placed in a 96 well plate (0.5 million cells per wellin 1% FBS medium) then a test compound of this invention was added atdifferent concentrations. After 10 minutes, the cells were activatedwith PHA (final concentration 2.5 μg/mL) and incubated for 20 hours at37° C. under CO₂. The final volume was 200 μL. Following incubation, thecells were centrifuged and the supernatants collected and stored at −70°C. prior to assaying for IL-2 production. A commercial ELISA kit (IL-2Eli-pair, Diaclone Research, Besancon, France) was used to detectproduction of IL-2, from which dose response curves were obtained. TheIC₅₀ value was calculated as the concentration at which 50% of maximumIL-2 production after stimulation was inhibited versus a non-stimulationcontrol.

Compound # IC₅₀ (nM) 1 3 2 7 3 8 4 15 5 10 6 2 7 3 8 6 9 9 10 31 11 1612 9 13 73 14 8 15 10 16 50 17 25 18 17 19 3 20 19 21 27 22 152 23 6 243 25 27

Inhibition of other cytokines, such as IL-4, IL-5, IL-13, GM-CSF, TNF-α,and INF-γ, can be tested in a similar manner using a commerciallyavailable ELISA kit for each cytokine.

Example B Patch Clamp Studies of Inhibition of I_(CRAC) Current in RBLCells, Jurkat Cells, and Primary T Cells

In general, a whole cell patch clamp method was used to examine theeffects of a compound of the invention on a channel that mediatesI_(crac). In such experiments, a baseline measurement was establishedfor a patched cell. Then a compound to be tested was perfused (orpuffed) to cells in the external solution and the effect of the compoundon I_(crac) was measured. A compound that modulates L_(crac) (e.g.,inhibits) is a compound that is useful in the invention for modulatingCRAC ion channel activity.

1) RBL Cells

Cells

Rat basophilic leukemia cells (RBL-2H3) were grown in DMEM mediasupplemented with 10% fetal bovine serum in an atmosphere of 95% air/5%CO₂. Cells were seeded on glass coverslips 1-3 days before use.

Recording Conditions

Membrane currents of individual cells were recorded using the whole-cellconfiguration of the patch clamp technique with an EPC10 (HEKAElectronik, Lambrecht, Germany). Electrodes (2-5 MΩ in resistance) werefashioned from borosilicate glass capillary tubes (Sutter Instruments,Novato, Ca). The recordings were done at room temperature.

Intracellular Pipette Solution

The intracellular pipette solution contained Cs-Glutamate 120 mM; CsC120mM; CsBAPTA 10 mM; CsHEPES 10 mM; NaCl 8 mM; MgCl₂ 1 mM; 1P3 0.02 mM;pH=7.4 adjusted with CsOH. The solution was kept on ice and shieldedfrom light before the experiment was preformed.

Extracellular Solution

The extracellular solution contained NaCl 138 mM; NaHEPES, 10 mM; CsCl10 mM; CaCl₂ 10 mM; Glucose 5.5 mM; KCl 5.4 mM; KH₂PO₄ 0.4 mM;Na₂HPO₄H₂O 0.3 mM at pH=7.4 adjusted with NaOH.

Compound Treatment

Each compound was diluted from a 10 mM stock in series using DMSO. Thefinal DMSO concentration was always kept at 0.1%.

Experimental Procedure

I_(CRAC) currents were monitored every 2 seconds using a 50 msecprotocol, where the voltage was ramped from −100 mV to +100 mV. Themembrane potential was held at 0 mV between the test ramps. In a typicalexperiment, the peak inward currents would develop within 50-100seconds. Once the I_(CRAC) currents were stabilized, the cells wereperfused with a test compound in the extracellular solution. At the endof an experiment, the remaining I_(CRAC) currents were then challengedwith a control compound (SKF96365, 10 μM) to ensure that the currentcould still be inhibited.

Data Analysis

The I_(CRAC) current level was determined by measuring the inwardcurrent amplitude at −80 mV of the voltage ramp in an off-line analysisusing MATLAB. The I_(CRAC) current inhibition for each concentration wascalculated using peak amplitude in the beginning of the experiment fromthe same cell. The IC₅₀ value and Hill coefficient for each compound wasestimated by fitting all the individual data points to a single Hillequation.

Results

The table below shows the concentration of compounds of the inventionwhich inhibits 50% of the I_(CRAC) current in RBL cells.

Compound Number IC₅₀  2 133 nM 24 147 nM SKF96365 4000 nM 

2) Jurkat Cells

Cells

Jurkat T cells are grown on glass coverslips, transferred to therecording chamber and kept in a standard modified Ringer's solution ofthe following composition: NaCl 145 mM, KCl 2.8 mM, CsCl 10 mM, CaCl₂ 10mM, MgCl₂ 2 mM, glucose 10 mM, HEPES.NaOH 10 mM, pH 7.2.

Extracellular Solution

The external solution contains 10 mM CaNaR, 11.5 mM glucose and a testcompound at various concentrations.

Intracellular Pipette Solution

The standard intracellular pipette solution contains: Cs-glutamate 145mM, NaCl 8 mM, MgCl₂ 1 mM, ATP 0.5 mM, GTP 0.3 mM, pH 7.2 adjusted withCsOH. The solution is supplemented with a mixture of 10 mM Cs-BAPTA and4.3-5.3 mM CaCl₂ to buffer [Ca²⁺]i to resting levels of 100-150 nM.

Patch-Clamp Recordings

Patch-clamp experiments are performed in the tight-seal whole-cellconfiguration at 21-25° C. High-resolution current recordings areacquired by a computer-based patch-clamp amplifier system (EPC-9, HEKA,Lambrecht, Germany). Sylgard®-coated patch pipettes typically haveresistances between 2-4 MΩ after filling with the standard intracellularsolution. Immediately following establishment of the whole-cellconfiguration, voltage ramps of 50 ms duration spanning the voltagerange of −100 to +100 mV are delivered from a holding potential of 0 mVat a rate of 0.5 Hz over a period of 300 to 400 seconds. All voltagesare corrected for a liquid junction potential of 10 mV between externaland internal solutions. Currents are filtered at 2.3 kHz and digitizedat 100 μs intervals. Capacitive currents and series resistance aredetermined and corrected before each voltage ramp using the automaticcapacitance compensation of the EPC-9.

Data Analysis

The very first ramps before activation of I_(CRAC) (usually 1 to 3) aredigitally filtered at 2 kHz, pooled and used for leak-subtraction of allsubsequent current records. The low-resolution temporal development ofinward currents is extracted from the leak-corrected individual rampcurrent records by measuring the current amplitude at −80 mV or avoltage of choice.

Compounds of the invention are expected to inhibit I_(CRAC) current inJurkat cells.

3) Primary T Cells

Preparation of Primary T Cells

Primary T cells are obtained from human whole blood samples by adding100 μL of RosetteSep® human T cell enrichment cocktail to 2 mL of wholeblood. The mixture is incubated for 20 minutes at room temperature, thendiluted with an equal volume of PBS containing 2% FBS. The mixture islayered on top of RosetteSep® DM-L density medium and then centrifugedfor 20 minutes at 1200 g at room temperature. The enriched T cells arerecovered from the plasma/density medium interface, then washed with PBScontaining 2% FBS twice, and used in patch clamp experiments followingthe procedure described for RBL cells.

Compounds of the invention are expected to inhibit I_(CRAC) current inhuman primary T cells.

Example C Inhibition of Multiple Cytokines in Primary Human PBMCs

Peripheral blood mononuclear cells (PBMCs) are stimulated withphytohemagglutinin (PHA) in the presence of varying concentrations ofcompounds of the invention or cyclosporine A (CsA), a known inhibitor ofcytokine production. Cytokine production is measured using commerciallyavailable human ELISA assay kits (from Cell Science, Inc.) following themanufacturers instructions.

The compounds of the invention are potent inhibitors of IL-2, and areexpected to be potent inhibitors of IL-4, IL-5, IL-13, GM-CSF, INF-α andTNF-γ in primary human PBM cells. In addition, compounds of theinvention are not expected to inhibit the anti-inflammatory cytokine,IL-10.

Example D Compounds of the Invention are Potent Inhibitors ofDegranulation in RBL Cells

Procedure:

The day before the assay is performed, RBL cells, that had been grown toconfluence in a 96 well plate, are incubated at 37° C. for at least 2hours. The medium is replaced in each well with 100 μL of fresh mediumcontaining 2 μLg/mL of anti-DNP IgE.

On the following day, the cells are washed once with PRS (2.6 mM glucoseand 0.1% BSA) and 160 μL of PRS was added to each well. A test compoundis added to a well in a 20 μL solution at 10× of the desiredconcentration and incubated for 20 to 40 minutes at 37° C. 20 μL of 10×mouse anti-IgE (10 μL/mL) is added. SKF96365 is used as a positivecontrol. Maximum degranulation typically occurs between 15 to 40 minutesafter addition of anti-IgE.

Results:

Compounds of the invention are expected to inhibit degranulation of RBLcells.

Example E Compounds of the Invention are Potent Inhibitors of Chemotaxisin T Cells

T-Cell Isolation:

Twenty ml aliquots of heparinized whole blood (2 pig, 1 human) aresubjected to density gradient centrifugation on Ficoll Hypaque. Thebuffy coat layers representing peripheral blood mononuclear cells(PBMCs) containing lymphocytes and monocytes are washed once,resuspended in 12 ml of incomplete RPMI 1640 and then placed ingelatin-coated T75 culture flasks for 1 hr at 37° C. The non-adherentcells, representing peripheral blood lymphocytes (PBLs) depleted ofmonocytes, are resuspended in complete RPMI media and placed in looselypacked activated nylon wool columns that had been equilibrated with warmmedia. After 1 hr at 37° C., the non-adherent T cell populations areeluted by washing of the columns with additional media. The T cellpreparations are centrifuged, resuspended in 5 ml of incomplete RPMI,and counted using a hemocytometer.

Cell Migration Assay:

Aliquots of each T cell preparation are labeled with Calcien AM(TefLabs) and suspended at a concentration of 2.4×10⁶/ml inHEPES-buffered Hank's Balanced Salt Solution containing 1.83 mM CaCl₂and 0.8 mM MgCl₂, pH 7.4 (HHBSS). An equal volume of HHBSS containing 0,20 nM, 200 nM or 2000 nM of compound 1 or 20 nM EDTA is then added andthe cells are incubated for 30 min at 37° C. Fifty μl aliquots of thecell suspensions (60,000 cells) are placed on the membrane (pore size 5μm) of a Neuroprobe ChemoTx 96 well chemotaxis unit that had beenaffixed over wells containing 10 ng/ml MIP-1α in HHBSS. The T cells areallowed to migrate for 2 hr at 37° C., after which the apical surface ofthe membrane is wiped clean of cells. The chemotaxis units are thenplaced in a CytoFlour 4000 (PerSeptive BioSystems) and the fluorescenceof each well is measured (excitation and emission wavelengths of 450 and530 nm, respectively). The number of migrating cells in each well isdetermined from a standard curve generated from measuring thefluorescence of serial two-fold dilutions of the labeled cells placed inthe lower wells of the chemotaxis unit prior to affixing the membrane.

Results:

Compounds of the invention are expected to be inhibitory to thechemotactic response of porcine T cells and in human T cells.

All publications, patent applications, patents, and other documentscited herein are incorporated by reference in their entirety. In case ofconflict, the present specification, including definitions, willcontrol. In addition, the materials, methods, and examples areillustrative only and not intended to be limiting in any way.

From the foregoing description, it will be apparent that variations andmodifications may be made to the invention described herein. Suchembodiments are also within the scope of the following claims.

I claim:
 1. A compound selected from the group consisting of:2,6-difluoro-N-(4-(5-(isoxazol-5-yl)-3-methylthiophen-2-yl)phenyl)benzamide;2,6-difluoro-N-(4-(3-methyl-5-(thiazol-2-yl)thiophen-2-yl)phenyl)benzamide;2,6-difluoro-N-(4-(3-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-2-yl)phenyl)benzamide;2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;2,6-difluoro-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;2,6-difluoro-N-(4-(3-methyl-5-(pyridin-3-yl)thiophen-2-yl)phenyl)benzamide;2,6-difluoro-N-(4-(2-methyl-5-(pyridin-3-yl)thiophen-3-yl)phenyl)benzamide;2,6-difluoro-N-(4-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)phenyl)benzamide;2,6-difluoro-N-(4-(2-methyl-5-(1,3,4-oxadiazol-2-yl)thiophen-3-yl)phenyl)benzamide;2,6-difluoro-N-(6-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-3-yl)benzamide;N-(4-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;2,6-difluoro-N-(4-(2-methyl-5-(4-methylpyridin-3-yl)thiophen-3-yl)phenyl)benzamide;N-(4-(5-(5-chloro-2-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyridin-2-yl)benzamide;2,6-difluoro-N-(5-(2-methyl-5-(thiazol-2-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyridin-2-yl)-2,6-difluorobenzamide;2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyridin-2-yl)benzamide;3-methyl-N-(5-(2-methyl-5-(oxazol-5-yl)thiophen-3-yl)pyridin-2-yl)isonicotinamide;N-(4-(5-(4-chloropyridin-3-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;2,6-difluoro-N-(4-(5-(4-methoxypyridin-3-yl)-2-methylthiophen-3-yl)phenyl)benzamide;2,6-difluoro-N-(5-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)pyrazin-2-yl)benzamide;N-(5-(5-(1H-imidazol-5-yl)-2-methylthiophen-3-yl)pyrazin-2-yl)-2,6-difluorobenzamide;2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamide;2,6-difluoro-N-(4-(2-methyl-5-(1-methyl-1H-imidazol-5-yl)thiophen-3-yl)phenyl)benzamidehydrochloride; andN-(4-(5-(1-ethyl-1H-imidazol-5-yl)-2-methylthiophen-3-yl)phenyl)-2,6-difluorobenzamide;or a pharmaceutically acceptable salt thereof.
 2. A pharmaceuticalcomposition, comprising a compound of claim 1, and: (i) apharmaceutically acceptable carrier; and/or (ii) one or more additionaltherapeutic agents.
 3. The pharmaceutical composition according to claim2, wherein the one or more additional therapeutic agents is selectedfrom the group consisting of immunosuppressive agents, anti-inflammatoryagents and mixtures thereof; or is selected from the group consisting ofsteroids, non-steroidal anti-inflammatory agents, antihistamines,analgesics, and suitable mixtures thereof.